QUICK SEARCH:   [advanced] Author: Keyword(s):
Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents

This Article Summary Figures Only Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Related articles in JCS Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Anderson, P. Articles by Kedersha, N. Search for Related Content PubMed PubMed Citation Articles by Anderson, P. Articles by Kedersha, N.

Stressful initiations

Division of Rheumatology, Immunology and Allergy, Brigham and Women's Hospital, Smith 652, One Jimmy Fund Way, Boston, MA 02115, USA

^* Author for correspondence (e-mail: panderson{at}rics.bwh.harvard.edu )

	Summary

Top Summary Introduction Stress granules: a historical... Stress granules: the mRNA... Stress granules: composition of... Stress granules and polysomes:... Translational triage model Translational silencing without... Nuclear history Conclusions/perspectives References

 
Stress granules (SGs) are phase-dense particles that appear in the cytoplasm of eukaryotic cells that have been exposed to environmental stress (e.g. heat, oxidative conditions, hyperosmolarity and UV irradiation). SG assembly is a consequence of abortive translational initiation: SGs appear when translation is initiated in the absence of eIF2-GTP-tRNA_i^Met, the ternary complex that normally loads tRNA_i^Met onto the small ribosomal subunit. Stress-induced depletion of eIF2-GTP-tRNA_i^Met allows the related RNA-binding proteins TIA-1 and TIAR to promote the assembly of eIF2-eIF5-deficient preinitiation complexes, the core constituents of SGs. The mRNP components that make up the SG are in a dynamic equilibrium with polysomes. As such, the SG appears to constitute a metabolic domain through which mRNPs are continually routed and subjected to triage — they are first monitored for integrity and composition, and then sorted for productive translational initiation or targeted degradation.

Key words: Stress granules, TIA-1, TIAR, Protein translation, eIF2alpha, Heat shock

	Introduction

Top Summary Introduction Stress granules: a historical... Stress granules: the mRNA... Stress granules: composition of... Stress granules and polysomes:... Translational triage model Translational silencing without... Nuclear history Conclusions/perspectives References

 
In response to environmental stress (e.g. heat, hyperosmolarity and oxidative conditions), eukaryotic cells shut down protein synthesis in a stereotypic response that conserves anabolic energy for the repair of stress-induced damage. The translational arrest that accompanies environmental stress is selective: whereas translation of constitutively expressed `housekeeping' transcripts is turned off, translation of stress-induced transcripts encoding heat shock proteins and some transcription factors is maintained or enhanced. The stress-activated signaling cascades responsible for reprogramming translation in stressed cells are becoming apparent. At the apex of these signaling cascades is a family of serine/threonine kinases that serve as sensors of environmental stress. Included in this family are the following: (1) PKR, a double-stranded RNA-dependent kinase that is activated by viral infection, heat and UV irradiation (Williams, 2001); (2) PERK/PEK, a resident ER protein that is activated when unfolded proteins accumulate in the ER (Harding et al., 2000; Patil and Walter, 2001); (3) GCN2, a protein that monitors amino acid levels in the cell and responds to amino acid deprivation (Kimball, 2001); and (4) HRI, a heme-regulated kinase that ensures the balanced synthesis of globin chains and heme during erythroctye maturation (Han et al., 2001; Lu et al., 2001). Each of these stress `sensors' phosphorylates eIF2, a critical regulatory component of the ternary complex (composed of eIF2ß bound to tRNA_i^Met and GTP) that loads initiator tRNA_i^Met onto the small ribosomal subunit to initiate protein synthesis (Dever, 2002; Kimball, 2001). Phosphorylation of eIF2 converts the eIF2 ternary complex into a competitive inhibitor of eIF2B, the GTP/GDP exchange factor that converts inactive ternary complex (GDP-associated) to active ternary complex (GTP-associated) (Krishnamoorthy et al., 2001). Thus, phosphorylation of eIF2 inhibits protein translation by reducing the availability of the eIF2-GTP-tRNA_i^Met ternary complex.

When translation is initiated in the absence of eIF2-GTP-tRNA_i^Met, an eIF2/eIF5-deficient, `stalled' 48S^* preinitiation complex is assembled (Kedersha et al., 2002). These eIF2/eIF5-deficient preinitiation complexes and their associated mRNA transcripts are dynamically routed to cytoplasmic foci known as stress granules (SGs), in a process that requires the related RNA-binding proteins TIA-1 and TIAR (Kedersha et al., 2002; Kedersha et al., 2000; Kedersha et al., 1999). In stressed cells, mRNA is in a dynamic equilibrium between polysomes and SGs (Kedersha et al., 2000). Here we discuss the antagonistic roles of eIF2 and TIA-1/TIAR in the assembly of polysomes and SGs in the context of a translational checkpoint model, wherein TIA and eIF2 are functional antagonists whose relative activities determine how many times a given transcript is translated before it is degraded.

	Stress granules: a historical perspective

Top Summary Introduction Stress granules: a historical... Stress granules: the mRNA... Stress granules: composition of... Stress granules and polysomes:... Translational triage model Translational silencing without... Nuclear history Conclusions/perspectives References

 
When exposed to supra-ambient temperatures, eukaryotic cells exhibit characteristic morphological changes in both nuclear and cytoplasmic compartments. In the nucleus, actin filaments condense to form rod-shaped bodies, and the granularity of nucleoli is altered (Collier et al., 1988; Collier and Schlesinger, 1986). In the cytoplasm, the cytoskeleton is rearranged and the Golgi apparatus is disrupted (Collier et al., 1988; Collier and Schlesinger, 1986). The appearance of phase-dense cytoplasmic granules was first noted in cultures of Peruvian tomato cells subjected to heat shock (Nover et al., 1983). Low-molecular-weight heat shock proteins were identified as prominent components of tomato heat shock granules (Nover et al., 1983). Immunofluorescence microscopy revealed that they are also components of phase-dense granules observed in cytoplasm of heat-shocked mammalian cells (Arrigo et al., 1988). The use of immunofluorescence microscopy to detect stress-induced changes in the subcellular localization of cellular components has allowed the identification of cytoplasmic and nuclear microdomains that are assembled as part of the normal stress response. Heat-shock-induced transcription factors have been found to accumulate at discrete cytoplasmic and nuclear foci in response to stress (Cotto et al., 1997; Cotto and Morimoto, 1999; Holmberg et al., 2000; Jolly et al., 2002; Scharf et al., 1998). Similarly, the subcellular localization of mRNP particles involved in various aspects of mRNA metabolism is altered in cells subjected to environmental stress (Gallouzi et al., 2000; Krebber et al., 1999; Michael et al., 1995; van der Houven van Oordt et al., 2000). Thus, immunofluorescence microscopy can be used to identify subcellular domains that may regulate cellular metabolism during stress.

	Stress granules: the mRNA connection

Top Summary Introduction Stress granules: a historical... Stress granules: the mRNA... Stress granules: composition of... Stress granules and polysomes:... Translational triage model Translational silencing without... Nuclear history Conclusions/perspectives References

 
The identification of mRNA as a component of plant heat shock granules provided a clue to the possible function of these cytoplasmic structures (Nover et al., 1989). Remarkably, heat-shock-granule-associated mRNAs were found to encode constitutively expressed `housekeeping' proteins but not newly synthesized heat shock proteins (Nover et al., 1989). This result suggested that translationally silenced mRNAs selectively accumulate at SGs, whereas translationally active mRNAs are excluded. These mRNAs could still be translated in vitro and could also be translated in vivo after the cells recovered from stress. Thus, the plant heat shock granule was proposed to serve as a storage repository for untranslated mRNAs.

Poly(A)⁺ RNA is also a component of mammalian SGs (Kedersha et al., 1999), as demonstrated by in situ hybridization using oligo-dT (Fig. 1). Recently, sequence-specific in situ hybridization has shown that mRNA encoding inducible HSP70 is selectively excluded from mammalian SGs (P.A. and N.K., unpublished). Thus, the SGs found in the cytoplasm of both plant and animal cells are sites at which untranslated mRNA accumulates in cells subjected to environmental stress.

View larger version (65K): [in this window] [in a new window]   Fig. 1. Assembly and disassembly of arsenite-induced stress granules. DU145 cells were cultured in the absence (control) or presence of arsenite (1 mM) for 30 minutes (STRESS), washed and allowed to recover for 1 hour or 3 hours (RECOVERY) before processing for two-color immunofluorescent microscopy. TIA-1 protein is identified using a polyclonal antibody (green). Poly(A)+ RNA is revealed by in situ hybridation using an oligo-dT probe (red). Sites of co-localization of TIA-1 and poly(A)+ RNA appear yellow. Nuclei are counterstained using Hoechst dye (blue).

	Stress granules: composition of the mRNP

Top Summary Introduction Stress granules: a historical... Stress granules: the mRNA... Stress granules: composition of... Stress granules and polysomes:... Translational triage model Translational silencing without... Nuclear history Conclusions/perspectives References

 
Identification of the protein components of mammalian SGs was facilitated by the discovery that the related RNA-binding proteins TIA-1 and TIAR are robust markers of these cytoplasmic foci (Kedersha et al., 1999). TIA-1 and TIAR are concentrated in the nucleus of most cells, but heterokaryon analysis reveals that both proteins continuously shuttle between the nucleus and the cytoplasm (P.A. and N.K., unpublished). In response to environmental stress, TIA-1 and TIAR accumulate in the cytoplasm, where they rapidly aggregate to form mammalian stress granules (Fig. 1) (Kedersha et al., 1999). Following removal of a non-lethal stress, the SGs increase in size owing to fusion of smaller SGs, and then rapidly disperse [Fig. 1; see movie of this process in Kedersha et al. (Kedersha et al., 2000)]. TIA-1 (Tian et al., 1991) and TIAR (Kawakami et al., 1992) possess three RNA-recognition motifs at their N-termini and a glutamine-rich domain at their C-termini. TIA mutants lacking the RNA-recognition motifs function as transdominant inibitors of SG assembly in stressed cells (Kedersha et al., 2000) which suggests that the TIA proteins are required for SG assembly.

The discovery of TIA-1 and TIAR as markers of SGs allowed the use of dual-labeling studies to identify additional SG components. This analysis revealed that components of small, but not large, ribosomal subunits co-localize with TIA-1 and TIAR at SGs (Kedersha et al., 2002). The absence of the large ribosomal subunit eliminated the possibility that SGs are sites at which selected mRNAs are translated during stress. At the same time, the presence of the small ribosomal subunit indicated that the mRNPs making up SGs might be related to polysomes. Consistent with this premise is compositional analysis revealing that many of the protein and RNA components of SGs and polysomes are identical (Kedersha et al., 2002).

Translation is normally initiated when the small ribosomal subunit and its associated initiation factors are recruited to a capped mRNA transcript to form a 48S complex (Fig. 2, left branch of pathway; green arrows). Hydrolysis of eIF2-associated GTP by eIF5 displaces the early initiation factors, allowing the binding of the large ribosomal subunit. Repeated cycles of successful initiation convert an mRNA into a polysome. In stressed cells, activation of one or more eIF2 kinases (e.g. PKR, PERK/PEK, GCN2, HRI; Fig. 2, red box) results in the phosphorylation of eIF2 (Kimball, 2001; Williams, 2001), which consequently inhibits eIF2B, the GTP/GDP exchange factor that charges the eIF2 ternary complex. The ensuing depletion of eIF2-GTP-tRNA_i^Met prevents productive translation initiation. Under these conditions, TIA-1 and TIAR promote the assembly of an eIF2/eIF5-deficient preinitiation complex (denoted 48S^* in Fig. 2) that is routed to SGs (Fig. 2, right branch of pathway, red arrows) (Kedersha et al., 2002). RNA-binding proteins that either promote [human autoantigen R (HuR) (Gallouzi et al., 2000)] or inhibit (tristetraprolin; P.A. and N.K., unpublished) mRNA stability are also recruited to SGs. This suggests that the SG is a site where the fates of specific transcripts are determined by the activity of different RNA-binding proteins. Whether the SG is also a site of mRNA processing (e.g. through degradation by exosomes) remains to be determined.

View larger version (37K): [in this window] [in a new window]   Fig. 2. Translational initiation in the absence or presence of stress. (Green panels) In the absence of stress, eIF2B promotes the charging of the eIF2-GTP-tRNAMet ternary complex by exchanging GDP for GTP. When the eIF2-GTP-tRNAMet ternary complex is available, a canonical 48S preinitiation complex is assembled at the 5' end of capped transcripts (green arrow: Normal) and scanning begins. Upon recognition of the initiation codon by the anticodon of tRNAMet, eIF5 promotes GTP hydrolysis, and early initiation factors are displaced by the 60S ribosomal subunit. As additional ribosomes are added to the transcript, the mRNA is converted into a polysome. (Red panels) In stressed cells (red arrow: Stress), phosphorylation of eIF2 by PKR, PERK, HRI or GCN2 converts eIF2 into a competitive antagonist of eIF2B, depleting the stores of eIF2/GTP/tRNAMet. Under these conditions, TIA-1 is included in a non-canonical, eIF2/eIF5-deficient 48S* preinitiation complex (composed of all components of the 48S pre-initiation complex except eIF2 and eIF5) that is translationally silent. TIA-1 self-aggregation then promotes the accumulation of these complexes at discrete cytoplasmic foci known as stress granules. Blue square, eIF5; green triangle, eIF2 bound to GTP; yellow triangle, eIF2 bound to GDP; red triangle, phospho-eIF2 bound to GDP.

	Stress granules and polysomes: a dynamic equilibrium

Top Summary Introduction Stress granules: a historical... Stress granules: the mRNA... Stress granules: composition of... Stress granules and polysomes:... Translational triage model Translational silencing without... Nuclear history Conclusions/perspectives References

 
Earlier models suggested that heat-shock-granules are storage depots for untranslated mRNAs, but mammalian SGs exhibit behavior inconsistent with such a model. The antagonistic effects of different pharmacological inhibitors of protein translation on SG assembly have revealed that SG-associated mRNA is in a dynamic equilibrium with polysomes (Fig. 3) (Kedersha et al., 2000). Drugs that stabilize polysomes by freezing ribosomes on translating mRNAs (e.g. cycloheximide and emetine) inhibit the assembly of SGs and actively dissolve SGs in the continued presence of both stress and eIF2 phosphorylation (Kedersha et al., 2000). Conversely, drugs that destabilize polysomes by releasing ribosomes from mRNA transcripts (e.g. puromycin), promote the assembly of SGs (Kedersha et al., 2000). This behavior suggested that SG-associated poly(A)+ RNA is in equilibrium with polysomes. Direct evidence for the dynamic nature of SGs was obtained in experiments using green fluorescent protein (GFP)-TIA-1 and a construct in which GFP was fused to poly(A)-binding protein (PABP-GFP) to monitor the assembly and disassembly of SGs in living cells (Kedersha et al., 2000). In response to arsenite-induced oxidative stress, GFP-TIA-1 rapidly moves from the nucleus to the cytoplasm, where it is evenly and diffusely distributed. After 10 minutes, the cytoplasmic GFP-TIA-1 aggregates into discrete foci that coalesce and slowly enlarge over the next 20 minutes. When arsenite is washed out of the cells, the SGs slowly disassemble with similar kinetics. However, this slow and steady accumulation of GFP-TIA-1 at SGs is misleading. Fluorescence recovery after photobleaching (FRAP) analysis reveals that GFP-TIA-1 shuttles in and out of SGs very rapidly such that 50% of SG-associated GFP-TIA-1 is replaced every 2 seconds. Although the rate at which mRNA shuttles in and out of SGs was not determined directly, GFP-PABP was used as a surrogate marker for its associated mRNA. Interestingly, GFP-PABP shuttles in and out of SGs at a rate that is ten times slower than that of GFP-TIA-1 (i.e. 50% of SG-associated GFP-PABP is replaced every 20 seconds). Given these kinetics, and considering that the dominant negative mutant of TIA-1 (e.g. TIA-1RRM) prevents SG assembly altogether (Kedersha et al., 2000), it appears that TIA-1 actively escorts untranslated mRNA to SGs. These data reveal that SGs are highly dynamic structures despite their apparent stability in real-time microscopy.

View larger version (26K): [in this window] [in a new window]   Fig. 3. The dynamic equilibrium between polysomes and stress granules is regulated by the availability of eIF2-GTP-tRNAMet and TIA proteins. Stress-induced activation of eIF2 kinases reduces the concentration of eIF2-GTP-tRNAMet, allowing TIA proteins to promote the assembly of stress granules. Several eukaryotic viruses [e.g. herpes simplex virus (HSV), adenovirus (adeno), vaccinia virus (VV) and influenza virus] prevent stress-induced translational arrest by inhibiting the activity of eIF2 kinases or activating eIF2 phosphatases. Green events and arrows indicate routes leading to translational activation; red events and arrows indicate events leading to translational inhibition.

The dynamic movement of mRNA into and out of SGs argues against a model in which SGs are passive repositories of untranslated mRNAs that accumulate in stressed cells. Rather, the SG is more likely to serve as a `way station' through which untranslated mRNAs pass before being translated or degraded (Kedersha et al., 2000). Taken together, the data suggest that TIA-1 and TIAR act downstream of the stress-induced phosphorylation of eIF2 to drive mRNA from polysomes to SGs (Fig. 3). The central importance of the PKR/eIF2 pathway in the cellular response to stress is indicated by the number of eukaryotic viruses that must disable PKR or reverse the phosphorylation of eIF2 to effect productive infection (Barber, 2001). If TIA-1 is an important component of the PKR/eIF2 stress response pathway, viruses might target TIA-1 to subvert this protective response. Indeed, TIA-1 and TIAR have been found to bind to specific sequences encoded by both West Nile Virus RNA (W. Li, Y. Li, N.K. et al., unpublished) and Sendai virus RNA (F. Iseni, D. Barcin, M. Nishhio et al., unpublished). Fig. 3 depicts the dynamic equilibrium between polysomes and SGs and summarizes the molecular pathways that regulate this equilibrium.

	Translational triage model

Top Summary Introduction Stress granules: a historical... Stress granules: the mRNA... Stress granules: composition of... Stress granules and polysomes:... Translational triage model Translational silencing without... Nuclear history Conclusions/perspectives References

 
Fig. 4 shows a model depicting the molecular stages in the conversion of a polysome into an SG and the subsequent fate of its component mRNAs. The formation of an eIF2-deficient 48S^* complex at the 5' end of the polysomal mRNA allows TIA to bind to RNA in lieu of ternary complex (stage 1), resulting in an abortive initiation event. This allows the previously initiated ribosomes to run off the transcript (stages 2 and 3) until only the 48S^* complex remains bound to the 5' end of the RNA (stage 4). Should an eIF5-eIF2-containing ternary complex become available at any time during this process, the transcript can still be reinitiated (green arrow) by displacement of TIA with eIF5-eIF2-GTP-tRNA_i^Met. However, as stress conditions disrupt TIA shuttling such that the normally nuclear TIA accumulates in the cytoplasm, more TIA becomes available to bind nonspecifically to the mRNA. The self-aggregation of TIA drives SG assembly (stages 5 and 6). Other RNA-binding proteins such as HuR and tristetraprolin (TTP) are recruited to the SG to determine the fate of specific transcripts. ATP depletion promotes the assembly of SGs without increasing phospho-eIF2 (Kedersha et al., 2002), and overexpression of HSP70 results in the dispersal of the prion-like domain of TIA-1 in vivo (P.A. and N.K., unpublished). Therefore, we propose that HSP70 and ATP are required to extricate mRNAs from the SG, probably by altering the conformation of the TIA prion-like domain. Thus, according to this model, phospho-eIF2 initiates the formation of SGs, a sudden flood of untranslated polyA(+) mRNA is released from polysomes, TIA proteins aggregate this RNA, and HSP70 (and ATP) is required to dis-aggregate the TIA proteins.

View larger version (44K): [in this window] [in a new window]   Fig. 4. Proposed mechanism for the assembly of stress granules. Assembly of an eIF2/eIF5-deficient preinitiation complex at the 5' end of a polysome results in translational arrest (1). As elongating ribosomes `run-off' the mRNA (2,3), the polysome is converted into a 48S* preinitiation complex (4) that is routed by TIA-1 into stress granules (5,6) or productively initiated by the replacement of TIA with eIF5/eIF2/GTP/tRNAmet (green circle). A requirement for HSP 70 and ATP in removing mRNAs from the SG is indicated. Destabilizing elements (red) such as tristetraprolin (TTP) are proposed to direct selected stress granule mRNAs to sites of degradation, whereas stabilizing elements such as HuR (blue) are proposed to direct selected mRNAs to sites of storage and/or reinitiation. By this triage process, the SG may monitor the structure and integrity of mRNP complexes and determine the fate of specific RNAs.

	Translational silencing without visible stress granules

Top Summary Introduction Stress granules: a historical... Stress granules: the mRNA... Stress granules: composition of... Stress granules and polysomes:... Translational triage model Translational silencing without... Nuclear history Conclusions/perspectives References

 
Overexpression of recombinant TIA-1 represses the production of co-expressed reporter genes in the absence of exogenous stress (Kedersha et al., 2000). It is therefore likely that some eIF2/eIF5-deficient `abortive' preinitiation complexes are assembled under normal conditions, and that their frequency determines how often mRNA transcripts are subjected to triage. Consistent with this view, endogenous TIA-1 and TIAR repress the translation of TNF transcripts in the absence of stress. Both TIA-1 and TIAR are components of a regulatory complex that binds to an adenine/uridine-rich element found in the 3' untranslated region of TNF transcripts (Gueydan et al., 1999; Piecyk et al., 2000). LPS-activated macrophages derived from mice lacking either TIA-1 or TIAR overexpress TNF compared with wild-type controls. In macrophages lacking TIA-1, the polysome profile of TNF transcripts is shifted such that the percentage of TNF transcripts associated with polysomes is increased compared with that of wild-type macrophages (Piecyk et al., 2000). This suggests that TIA-1 represses the translation of TNF by promoting the assembly of non-polysomal mRNP complexes. Sucrose-density-gradient-analysis reveals that TIA-1 is found in low-density fractions that contain soluble proteins, as well as higher density fractions that contain 40-60S mRNPs, but not in polysomes (Kedersha et al., 2000). It is likely that the TIA-mediated shift of TNF- mRNA transcripts away from polysomes occurs through the assembly of abortive eIF2/eIF5-deficient pre-initiation complexes that are similar or identical to those that comprise the core units of SGs.

	Nuclear history

Top Summary Introduction Stress granules: a historical... Stress granules: the mRNA... Stress granules: composition of... Stress granules and polysomes:... Translational triage model Translational silencing without... Nuclear history Conclusions/perspectives References

 
Their visible roles in the cytoplasm notwithstanding, TIA-1 and TIAR normally predominate in the nucleus, where they have been shown to act as selective regulators of alternative mRNA splicing (Del Gatto-Konczak et al., 2000; Forch et al., 2000; Le Guiner et al., 2001). The binding of either TIA-1 or TIAR to uridine-rich elements found in intronic sequences located downstream of weak 5' splice sites promotes the recruitment of U1snRNP and the inclusion of `cryptic' alternative exons that are otherwise excised from the heteronuclear RNA. In their dual ability to regulate both mRNA splicing and translation, TIA-1 and TIAR resemble several other multifunctional RNA-binding proteins, including PTB, CUB-BP-related proteins, La, hnRNPK and hnRNP A1 (Wilkinson and Shyu, 2001; Ladomery, 1997). The process of mRNA splicing has been shown to `mark' selected mRNA transcripts for both quality control in the nucleus and translational control in the cytoplasm (Le Hir et al., 2000a; Le Hir et al., 2000b). It is therefore possible that heteronuclear mRNAs encoding introns that are recognized by TIA-1/TIAR retain these proteins at the exon-exon junction following the removal of the intron upon splicing. Upon arrival in the cytoplasm, transcripts that are `marked' by TIA-1/TIAR could be selectively regulated at the level of mRNA stability or translatability. Such an explanation would allow for the nuclear loading of TIA-1 and TIAR onto selected transcripts that become subject to translational silencing once in the cytoplasm. It also suggests a mechanism whereby stress-induced transcripts might be excluded from SGs: mRNAs transcribed during stress would escape being marked by TIA proteins, whose normal shuttling appears to be disrupted when they are routed into cytoplasmic SGs.

	Conclusions/perspectives

Top Summary Introduction Stress granules: a historical... Stress granules: the mRNA... Stress granules: composition of... Stress granules and polysomes:... Translational triage model Translational silencing without... Nuclear history Conclusions/perspectives References

 
Molecular modifications (particularly phosphorylation) are widely appreciated to regulate many distinct steps of the translational initiation process (Clemens, 2001; Lawrence and Brunn, 2001; Mahalingam and Cooper, 2001). The importance of subcellular compartmentalization of translational components is an important mode of regulation that occurs in living cells but is less understood. Key components of the translational apparatus (e.g. mRNA and its associated proteins, ribosomal subunits and translation initiation factors) move between the nucleus and the cytoplasm in a regulated manner. The localized availability and hence the activity of these components is regulated by interactions with organelles and with the cytoskeleton. The rapid assembly and disassembly of mammalian SGs is a striking illustration of this type of regulation. SGs coalesce into existence as mRNAs are released from polysomes during stress-induced translational arrest and melt away like snowflakes as the cell adapts or recovers from the stress.

The molecular trigger for SG formation is the abortive translation that occurs when a transcript is initiated without the eIF2-GTP-tRNA^Met ternary complex. The assembly of translationally inactive initiation complexes lacking eIF2, coupled with increased levels of cytoplasmic TIA, allows the RNA-binding proteins TIA-1 and/or TIAR to redirect untranslated mRNAs from polyribosomes to SGs (Fig. 4). Thus, TIA-1/TIAR and eIF2-GTP-tRNA_i^Met appear to act as antagonists that regulate the equilibrium between polysomes and SGs (Figs 3, 4). It remains to be determined whether TIA-1 and eIF2-GTP-tRNA^Met compete for binding to a common site on the preinitiation complex. Regardless of the details of molecular mechanism, the functional antagonism between eIF2 and TIA may determine the frequency with which a given mRNA transcript is initiated before being subject to a checkpoint at which mRNP structure and composition is monitored. If the ratio of TIA-1/TIAR to eIF2-GTP-tRNA^Met were 1:10, one might predict that, on average, ten productive initiation events would occur before a TIA-1/TIAR-containing translationally incompetent pre-initiation complex is assembled. As translating ribosomes `run off' this mRNA, the eIF2/eIF5-deficient complex would be directed to an SG, where the integrity and composition of the mRNP might determine whether the transcript is reinitiated, degraded or packaged into an untranslated mRNP particle.

Although considerable progress has been made in elucidating the relationship between SGs and translational initiation, we have much to learn about the connection between SGs and molecular chaperones. The aggregation domain of TIA is its glutamine-rich C-terminus, which resembles prion protein, and is essential for SG assembly. Specific molecular chaperones are required to disperse aggregated TIA-1 in living cells (P.A. and N.K., unpublished). These chaperones are also required for the adaptive response to stress, which suggests that SGs constitute a point of crosstalk between molecular chaperones and eIF2 kinases. Do SGs function as signal transduction domains? And what else is in SGs? Plant SGs contain not only RNA and small HSPs but also heat shock transcription factors. Do mammalian SGs also contain stress-related transcription factors and, if so, which ones? The end of stress is not yet in sight!

	Footnotes

 
eIF, eukaryotic initiation factor; GCN2, general control nonrepressed 2; HRI, heme-regulated inhibitor; PERK, PKR-like endoplasmic reticulum kinase; PKR, double-stranded RNA-dependent protein kinase; RRM, RNA-recognition motif; TIA-1, T-cell intracellular antigen-1; TIAR, T-cell intracellular antigen-related.

	References

Top Summary Introduction Stress granules: a historical... Stress granules: the mRNA... Stress granules: composition of... Stress granules and polysomes:... Translational triage model Translational silencing without... Nuclear history Conclusions/perspectives References

 

Arrigo, A. P., Suhan, J. P. and Welch, W. J. (1988). Dynamic changes in the structure and intracellular locale of the mammalian low-molecular-weight heat shock protein. Mol. Cell. Biol. 8,5059 -5071.[Abstract/Free Full Text]

Barber, G. N. (2001). Host defense, viruses and apoptosis. Cell Death Differ. 8, 113-126.[Medline]

Clemens, M. J. (2001). Initiation factor eIF2 alpha phosphorylation in stress responses and apoptosis. Prog. Mol. Subcell. Biol. 27,57 -89.[Medline]

Collier, N. C. and Schlesinger, M. J. (1986). The dynamic state of heat shock proteins in chicken embryo fibroblasts. J. Cell Biol. 103,1495 -1507.[Abstract/Free Full Text]

Collier, N. C., Heuser, J., Levy, M. A. and Schlesinger, M. J. (1988). Ultrastructural and biochemical analysis of the stress granule in chicken embryo fibroblasts. J. Cell Biol. 106,1131 -1139.[Abstract/Free Full Text]

Cotto, J. J. and Morimoto, R. I. (1999). Stress-induced activation of the heat-shock response: cell and molecular biology of heat-shock factors. Biochem. Soc. Symp. 64,105 -118.[Medline]

Cotto, J., Fox, S. and Morimoto, R. (1997). HSF1 granules: a novel stress-induced nuclear compartment of human cells. J. Cell Sci. 110,2925 -2934.[Abstract]

Del Gatto-Konczak, F., Bourgeois, C. F., le Guiner, C., Kister, L., Gesnel, M. C., Stevenin, J. and Breathnach, R. (2000). The RNA-binding protein TIA-1 is a novel mammalian splicing regulator acting through intron sequences adjacent to a 5' splice site. Mol. Cell. Biol. 20,6287 -6299.[Abstract/Free Full Text]

Dever, T. E. (2002). Gene-specific regulation by general translation factors. Cell 108,545 -556.[Medline]

Forch, P., Puig, O., Kedersha, N., Martinez, C., Granneman, S., Seraphin, B., Anderson, P. and Valcarcel, J. (2000). The apoptosis-promoting factor TIA-1 is a regulator of alternative pre- mRNA splicing. Mol. Cell 6,1089 -1098.[Medline]

Gallouzi, I. E., Brennan, C. M., Stenberg, M. G., Swanson, M. S., Eversole, A., Maizels, N. and Steitz, J. A. (2000). HuR binding to cytoplasmic mRNA is perturbed by heat shock. Proc. Natl. Acad. Sci. USA 97,3073 -3078.[Abstract/Free Full Text]

Gueydan, C., Droogmans, L., Chalon, P., Huez, G., Caput, D. and Kruys, V. (1999). Identification of TIAR as a protein binding to the translational regulatory AU-rich element of tumor necrosis factor mRNA. J. Biol. Chem. 274,2322 -2326.[Abstract/Free Full Text]

Han, A. P., Yu, C., Lu, L., Fujiwara, Y., Browne, C., Chin, G., Fleming, M., Leboulch, P., Orkin, S. H. and Chen, J. J. (2001). Heme-regulated eIF2alpha kinase (HRI) is required for translational regulation and survival of erythroid precursors in iron deficiency. EMBO J. 20,6909 -6918.[Medline]

Harding, H. P., Novoa, I., Zhang, Y., Zeng, H., Wek, R., Schapira, M. and Ron, D. (2000). Regulated translation initiation controls stress-induced gene expression in mammalian cells. Mol. Cell 6,1099 -1108.[Medline]

Holmberg, C. I., Illman, S. A., Kallio, M., Mikhailov, A. and Sistonen, L. (2000). Formation of nuclear HSF1 granules varies depending on stress stimuli. Cell Stress Chaperones 5,219 -228.[Medline]

Jolly, C., Konecny, L., Grady, D. L., Kutskova, Y. A., Cotto, J. J., Morimoto, R. I. and Vourc'h, C. (2002). In vivo binding of active heat shock transcription factor 1 to human chromosome 9 heterochromatin during stress. J. Cell Biol. 156,775 -781.[Abstract/Free Full Text]

Kawakami, A., Tian, Q., Duan, X., Streuli, M., Schlossman, S. F. and Anderson, P (1992). Identification and functional characterization of a TIA-1-related nucleolysin. Proc. Natl. Acad. Sci. USA 89,8681 -8685.[Abstract/Free Full Text]

Kedersha, N. L., Gupta, M., Li, W., Miller, I. and Anderson, P. (1999). RNA-binding proteins TIA-1 and TIAR link the phosphorylation of eIF-2 alpha to the assembly of mammalian stress granules. J. Cell Biol. 147,1431 -1442.[Abstract/Free Full Text]

Kedersha, N., Cho, M., Li, W., Yacono, P., Chen, S., Gilks, N., Golan, D. and Anderson, P. (2000). Dynamic shuttling of TIA-1 accompanies the recruitment of mRNA to mammalian stress granules. J. Cell Biol. 151,1257 -1268.[Abstract/Free Full Text]

Kedersha, N., Chen, S., Gilks, N., Li, W., Miller, I., Stahl, J. and Anderson, P. (2002). Evidence that ternary complex (eIF2-GTP-tRNAMet)-deficient preinitiation complexes are core constituents of mammalian stress granules. Mol. Biol. Cell 13,195 -210.[Abstract/Free Full Text]

Kimball, S. R. (2001). Regulation of translation initiation by amino acids in eukaryotic cells. Prog. Mol. Subcell. Biol. 26,155 -184.[Medline]

Krebber, H., Taura, T., Lee, M. S. and Silver, P. A. (1999). Uncoupling of the hnRNP Np13p from mRNAs during the stress-induced block in mRNA export. Genes Dev 13,1994 -2004.[Abstract/Free Full Text]

Krishnamoorthy, T., Pavitt, G. D., Zhang, F., Dever, T. E. and Hinnebusch, A. G. (2001). Tight binding of the phosphorylated alpha subunit of initiation factor 2 (eIF2alpha) to the regulatory subunits of guanine nucleotide exchange factor eIF2B is required for inhibition of translation initiation. Mol. Cell. Biol. 21,5018 -5030.[Abstract/Free Full Text]

Ladomery, M. (1997). Multifunctional proteins suggest connections between transcriptional and post-transcriptional processes. Bioessays 19,903 -909.[Medline]

Lawrence, J. C., Jr and Brunn, G. J. (2001). Insulin signaling and the control of PHAS-I phosphorylation. Prog. Mol. Subcell. Biol. 26,1 -31.[Medline]

Le Guiner, C., Lejeune, F., Galiana, D., Kister, L., Breathnach, R., Stevenin, J. and del Gatto-Konczak, F. (2001). TIA-1 and TIAR activate splicing of alternative exons with weak 5' splice sites followed by a U-rich stretch on their own pre-mRNAs. J. Biol. Chem. 276,40638 -40646.[Abstract/Free Full Text]

Le Hir, H., Izaurralde, E., Maquat, L. E. and Moore, M. J. (2000a). The spliceosome deposits multiple proteins 20-24 nucleotides upstream of mRNA exon-exon junctions. EMBO J. 19,6860 -6869.[Medline]

Le Hir, H., Moore, M. J. and Maquat, L. E. (2000b). Pre-mRNA splicing alters mRNP composition: evidence for stable association of proteins at exon-exon junctions. Genes Dev. 14,1098 -1108.[Abstract/Free Full Text]

Lu, L., Han, A. P. and Chen, J. J. (2001). Translation initiation control by heme-regulated eukaryotic initiation factor 2alpha kinase in erythroid cells under cytoplasmic stresses. Mol. Cell. Biol. 21,7971 -7980.[Abstract/Free Full Text]

Mahalingam, M. and Cooper, J. A. (2001). Phosphorylation of mammalian eIF4E by Mnk1 and Mnk2: tantalizing prospects for a role in translation. Prog. Mol. Subcell. Biol. 27,132 -142.[Medline]

Michael, M. W., Choi, M. and Dreyfuss, G. (1995). A nuclear export signal in hnRNP A1: A signal-mediated, temperature-dependent nuclear protein export pathway. Cell 83,415 -422.[Medline]

Nover, L., Scharf, K. D. and Neumann, D. (1983). Formation of cytoplasmic heat shock granules in tomato cell cultures and leaves. Mol. Cell. Biol. 3,1648 -1655.[Abstract/Free Full Text]

Nover, L., Scharf, K. and Neumann, D. (1989). Cytoplasmic heat shock granules are formed from precursor particles and are associated with a specific set of mRNAs. Mol. Cell. Biol. 9,1298 -1308.[Abstract/Free Full Text]

Patil, C. and Walter, P. (2001). Intracellular signaling from the endoplasmic reticulum to the nucleus: the unfolded protein response in yeast and mammals. Curr. Opin. Cell Biol. 13,349 -355.[Medline]

Piecyk, M., Wax, S., Beck, A., Kedersha, N., Gupta, M., Maritim, B., Chen, S., Gueydan, C., Kruys, V., Streuli, M. and Anderson, P. (2000). TIA-1 is a translational silencer that selectively regulates the expression of TNF-alpha. EMBO J. 19,4154 -4163.[Medline]

Scharf, K. D., Heider, H., Hohfeld, I., Lyck, R., Schmidt, E. and Nover, L. (1998). The tomato Hsf system: HsfA2 needs interaction with HsfA1 for efficient nuclear import and may be localized in cytoplasmic heat stress granules. Mol. Cell. Biol. 18,2240 -2251.[Abstract/Free Full Text]

Tian, Q., Streuli, M., Saito, H., Schlossman, S. F. and Anderson, P. (1991). A polyadenylate binding protein localized to the granules of cytolytic lymphocytes induces DNA fragmentation in target cells. Cell 67,629 -639.[Medline]

van der Houven van Oordt, W., Diaz-Meco, M. T., Lozano, J., Krainer, A. R., Moscat, J. and Caceres, J. F. (2000). The MKK(3/6)-p38-signaling cascade alters the subcellular distribution of hnRNP A1 and modulates alternative splicing regulation. J. Cell Biol. 149,307 -316.[Abstract/Free Full Text]

Wilkinson, M. F. and Shyu, A. B. (2001). Multifunctional regulatory proteins that control gene expression in both the nucleus and the cytoplasm. Bioessays 23,775 -787.[Medline]

Williams, B. R. (2001). Signal integration via PKR. Sci. STKE 2001,RE2 .

Related articles in JCS:

Stress granules: way stations for mRNA

JCS 2002 115: 1601. [Full Text]  

This article has been cited by other articles:

				H. H. Kim, K. Abdelmohsen, A. Lal, R. Pullmann Jr., X. Yang, S. Galban, S. Srikantan, J. L. Martindale, J. Blethrow, K. M. Shokat, et al. Nuclear HuR accumulation through phosphorylation by Cdk1 Genes & Dev., July 1, 2008; 22(13): 1804 - 1815. [Abstract] [Full Text] [PDF]

				H. Montero, M. Rojas, C. F. Arias, and S. Lopez Rotavirus Infection Induces the Phosphorylation of eIF2{alpha} but Prevents the Formation of Stress Granules J. Virol., February 1, 2008; 82(3): 1496 - 1504. [Abstract] [Full Text] [PDF]

				J.-E. Kim, I. Ryu, W. J. Kim, O.-K. Song, J. Ryu, M. Y. Kwon, J. H. Kim, and S. K. Jang Proline-Rich Transcript in Brain Protein Induces Stress Granule Formation Mol. Cell. Biol., January 15, 2008; 28(2): 803 - 813. [Abstract] [Full Text] [PDF]

				H. Lelouard, E. K. Schmidt, V. Camosseto, G. Clavarino, M. Ceppi, H.-T. Hsu, and P. Pierre Regulation of translation is required for dendritic cell function and survival during activation J. Cell Biol., December 31, 2007; 179(7): 1427 - 1439. [Abstract] [Full Text] [PDF]

				S. Kwon, Y. Zhang, and P. Matthias The deacetylase HDAC6 is a novel critical component of stress granules involved in the stress response Genes & Dev., December 15, 2007; 21(24): 3381 - 3394. [Abstract] [Full Text] [PDF]

				S. Yamasaki, G. Stoecklin, N. Kedersha, M. Simarro, and P. Anderson T-cell Intracellular Antigen-1 (TIA-1)-induced Translational Silencing Promotes the Decay of Selected mRNAs J. Biol. Chem., October 12, 2007; 282(41): 30070 - 30077. [Abstract] [Full Text] [PDF]

				H. S. Kim, Y. Kuwano, M. Zhan, R. Pullmann Jr., K. Mazan-Mamczarz, H. Li, N. Kedersha, P. Anderson, M. C. J. Wilce, M. Gorospe, et al. Elucidation of a C-Rich Signature Motif in Target mRNAs of RNA-Binding Protein TIAR Mol. Cell. Biol., October 1, 2007; 27(19): 6806 - 6817. [Abstract] [Full Text] [PDF]

				R. Pullmann Jr., H. H. Kim, K. Abdelmohsen, A. Lal, J. L. Martindale, X. Yang, and M. Gorospe Analysis of Turnover and Translation Regulatory RNA-Binding Protein Expression through Binding to Cognate mRNAs Mol. Cell. Biol., September 15, 2007; 27(18): 6265 - 6278. [Abstract] [Full Text] [PDF]

				M. M. Emara and M. A. Brinton Interaction of TIA-1/TIAR with West Nile and dengue virus products in infected cells interferes with stress granule formation and processing body assembly PNAS, May 22, 2007; 104(21): 9041 - 9046. [Abstract] [Full Text] [PDF]

				T. Rzymski and A. L. Harris The Unfolded Protein Response and Integrated Stress Response to Anoxia Clin. Cancer Res., May 1, 2007; 13(9): 2537 - 2540. [Abstract] [Full Text] [PDF]

				U. Nonhoff, M. Ralser, F. Welzel, I. Piccini, D. Balzereit, M.-L. Yaspo, H. Lehrach, and S. Krobitsch Ataxin-2 Interacts with the DEAD/H-Box RNA Helicase DDX6 and Interferes with P-Bodies and Stress Granules Mol. Biol. Cell, April 1, 2007; 18(4): 1385 - 1396. [Abstract] [Full Text] [PDF]

				M. B. Decca, M. A. Carpio, C. Bosc, M. R. Galiano, D. Job, A. Andrieux, and M. E. Hallak Post-translational Arginylation of Calreticulin: A NEW ISOSPECIES OF CALRETICULIN COMPONENT OF STRESS GRANULES J. Biol. Chem., March 16, 2007; 282(11): 8237 - 8245. [Abstract] [Full Text] [PDF]

				K. A. Suprenant, N. Bloom, J. Fang, and G. Lushington The major vault protein is related to the toxic anion resistance protein (TelA) family J. Exp. Biol., March 15, 2007; 210(6): 946 - 955. [Abstract] [Full Text] [PDF]

				S. Gallois-Montbrun, B. Kramer, C. M. Swanson, H. Byers, S. Lynham, M. Ward, and M. H. Malim Antiviral Protein APOBEC3G Localizes to Ribonucleoprotein Complexes Found in P Bodies and Stress Granules J. Virol., March 1, 2007; 81(5): 2165 - 2178. [Abstract] [Full Text] [PDF]

				V. Stribinskis and K. S. Ramos Rpm2p, a protein subunit of mitochondrial RNase P, physically and genetically interacts with cytoplasmic processing bodies Nucleic Acids Res., February 28, 2007; 35(4): 1301 - 1311. [Abstract] [Full Text] [PDF]

K. Thama