Advertisement
Journal Article
All three PACSIN isoforms bind to endocytic proteins and inhibit endocytosis
J. Modregger, B. Ritter, B. Witter, M. Paulsson, M. Plomann
Journal of Cell Science 2000 113: 4511-4521;

Summary

The PACSINs are a family of cytoplasmic phosphoproteins that play a role in vesicle formation and transport. We report the cloning and cDNA sequencing of PACSIN 3 and the analysis of all three PACSIN isoforms with regard to tissue distribution, ligand binding properties and influence on endocytosis. PACSIN 3 differs from the other family members in having a short proline-rich region and lacking asparagine-proline-phenylalanine motifs. In contrast to the neurospecific PACSIN 1 and the ubiquitously expressed PACSIN 2, PACSIN 3 is mainly detected in lung and muscle tissues. All isoforms potentially oligomerize and bind to dynamin, synaptojanin 1 and N-WASP via their Src homology 3 domains. The PACSIN proteins colocalize with dynamin, but not with clathrin, implying a specific role with a distinct subpopulation of dynamin at defined cellular sites. Transferrin endocytosis is blocked in a dose-dependent manner in cells overexpressing the PACSIN variants, but the inhibitory effect can be abolished by mutating specific amino acid residues in the Src homology 3 domains. These characteristics of the PACSIN protein family suggest a general function in recruitment of the interacting proteins to sites of endocytosis.

REFERENCES

    1. Altschul, S. F.,
    2. Madden, T. L.,
    3. Schaffer, A. A.,
    4. Zhang, J.,
    5. Zhang, Z.,
    6. Miller, W. and
    7. Lipman, D. J.
    (1997). Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25, 3389–3402
    OpenUrlAbstract/FREE Full Text
    1. Benmerah, A.,
    2. Begue, B.,
    3. Dautry-Varsat, A. and
    4. Cerf-Bensussan, N.
    (1996). The ear of-adaptin interacts with the COOH-terminal domain of the Eps15 protein. J. Biol. Chem 271, 12111–12116
    OpenUrlAbstract/FREE Full Text
    1. Berger, B.,
    2. Wilson, D. B.,
    3. Wolf, E.,
    4. Tonchev, T.,
    5. Milla, M. and
    6. Kim, P. S.
    (1995). Predicting coiled coils by use of pairwise residue correlations. Proc. Natl. Acad. Sci. USA 92, 8259–8263
    OpenUrlAbstract/FREE Full Text
    1. Cao, H.,
    2. Garcia, F. and
    3. McNiven, M. A.
    (1998). Differential distribution of dynamin isoforms in mammalian cells. Mol. Biol. Cell 9, 2595–2609
    OpenUrlAbstract/FREE Full Text
    1. Cook, T.,
    2. Mesa, K. and
    3. Urrutia, R.
    (1996). Three dynamin-encoding genes are differentially expressed in developing rat brain. J. Neurochem 67, 927–931
    OpenUrlCrossRefPubMedWeb of Science
    1. Cremona, O.,
    2. Di Paolo, G.,
    3. Wenk, M. R.,
    4. Luthi, A.,
    5. Kim, W. T.,
    6. Takei, K.,
    7. Daniell, L.,
    8. Nemoto, Y.,
    9. Shears, S. B.,
    10. Flavell, R. A. and
    11. et al
    . (1999). Essential Role of Phosphoinisitide Metabolism in Synaptic Vesicle Recycling. Cell 99, 179–188
    OpenUrlCrossRefPubMedWeb of Science
    1. Damke, H.,
    2. Baba, T.,
    3. Warnock, D. E. and
    4. Schmid, S. L.
    (1994). Induction of mutant dynamin specifically blocks endocytic coated vesicle formation. J. Cell Biol 127, 915–934
    OpenUrlAbstract/FREE Full Text
    1. Dowbenko, D.,
    2. Spencer, S.,
    3. Quan, C. and
    4. Lasky, L. A.
    (1998). Identification of a novel polyproline recognition site in the cytoskeletal associated protein, proline serine threonine phosphatase interacting protein. J. Biol. Chem 273, 989–996
    OpenUrlAbstract/FREE Full Text
    1. Fazioli, F.,
    2. Minichiello, L.,
    3. Matoskova, B.,
    4. Wong, W. T. and
    5. Di Fiore, P. P.
    ) (1993). Eps15, a novel tyrosine kinase substrate, exhibits transforming activity. Mol. Cell Biol 13, 5814–5828
    OpenUrlAbstract/FREE Full Text
    1. Giachino, C.,
    2. Lantelme, E.,
    3. Lanzetti, L.,
    4. Saccone, S.,
    5. Della Valle, G. and
    6. Migone, N.
    (1997). A novel SH3-containing human gene family preferentially expressed in the central nervous system. Genomics 41, 427–434
    OpenUrlCrossRefPubMedWeb of Science
    1. Greengard, P.,
    2. Valtorta, F.,
    3. Czernik, A. J. and
    4. Benfenati, F.
    (1993). Synaptic vesicle phosphoproteins and regulation of synaptic function. Science 259, 780–785
    OpenUrlAbstract/FREE Full Text
    1. Guo, S.,
    2. Stolz, L. E.,
    3. Lemrow, S. M. and
    4. York, J. D.
    (1999). SAC1 -like domains of yeast SAC1, INP52 and INP53 and of human synaptojanin encode polyphosphoinositide phosphatases. J. Biol. Chem 274, 12990–12995
    OpenUrlAbstract/FREE Full Text
    1. Henley, J. R. and
    2. McNiven, M. A.
    (1996). Association of a dynamin-like protein with the Golgi apparatus in mammalian cells. J. Cell Biol 133, 761–775
    OpenUrlAbstract/FREE Full Text
    1. Hinshaw, J. E. and
    2. Schmid, S. L.
    (1995). Dynamin self-assembles into rings suggesting a mechanism for coated vesicle budding. Nature 374, 190–192
    OpenUrlCrossRefPubMedWeb of Science
    1. Howard, L.,
    2. Nelson, K. K.,
    3. Maciewicz, R. A. and
    4. Blobel, C. P.
    (1999). Interaction of the Metalloprotease Disintegrins MDC9 and MDC15 with two SH3 domain-containing proteins, Endophilin I and SH3PX1. J. Biol. Chem 274, 31693–31699
    OpenUrlAbstract/FREE Full Text
    1. Kozak, M.
    (1991). An analysis of vertebrate mRNA sequences: Intimations of translational control. J. Cell Biol 115, 887–903
    OpenUrlAbstract/FREE Full Text
    1. Le Borgne, R. and
    2. Hoflack, B.
    (1998). Mechanisms of protein sorting and coat assembly: insights from the clathrin-coated vesicle pathway. Curr. Opin. Cell Biol 10, 499–503
    OpenUrlCrossRefPubMedWeb of Science
    1. Li, J.,
    2. Nishizawa, K.,
    3. An, W.,
    4. Hussey, R. E.,
    5. Lialios, F. E.,
    6. Salgia, R.,
    7. Sunder-Plassmann, R. and
    8. Reinherz, E. I.
    (1998). A cdc15-like adaptor protein (CD2BP1) interacts with the CD2 cytoplasmic domain and regulates CD2-triggered adhesion. EMBO J 17, 7320–7336
    OpenUrlAbstract
    1. Lupas, A.,
    2. Van Dyke, M. and
    3. Stock, J.
    (1991). Predicting coiled coils from protein sequences. Science 252, 1162–1164
    OpenUrlFREE Full Text
    1. McPherson, P. S.,
    2. Garcia, E. P.,
    3. Slepnev, V. I.,
    4. David, C.,
    5. Zhang, X.,
    6. Grabs, D.,
    7. Sossin, W. S.,
    8. Bauerfeind, R.,
    9. Nemoto, Y. and
    10. De Camilli, P.
    (1996). A presynaptic inositol-5-phosphatase. Nature 379, 353–356
    OpenUrlCrossRefPubMedWeb of Science
    1. Meriläinen, J.,
    2. Lehto, V.-P. and
    3. Wasenius, V.-M.
    ) (1997). FAP52, a novel, SH3 domain-containing focal adhesion protein. J. Biol. Chem 272, 23278–23284
    OpenUrlAbstract/FREE Full Text
    1. Miki, H.,
    2. Miura, K. and
    3. Takenawa, T.
    (1996). N-WASP, a novel actin-depolymerizing protein, regulates the cortical cytoskeletal rearrangement in PIP2-dependent manner downstream of tyrosine kinases. EMBO J 15, 5326–5335
    OpenUrlPubMedWeb of Science
    1. Miki, H.,
    2. Sasaki, T.,
    3. Takai, Y. and
    4. Takenawa, T.
    (1998). Induction of filopodium formation by a WASP-related actin-depolymerizing protein N-WASP. Nature 391, 93–96
    OpenUrlCrossRefPubMed
    1. Paoluzi, S.,
    2. Castagnoli, L.,
    3. Lauro, I.,
    4. Salcini, A. E.,
    5. Coda, L.,
    6. Fre, S.,
    7. Confalonieri, S.,
    8. Pelicci, P. G.,
    9. Di Fiore, P. P. and
    10. Cesareni, G.
    (1998). Recognition specificity of individual EH domains of mammals and yeast. EMBO J 17, 6541–6550
    OpenUrlAbstract
    1. Plomann, M.,
    2. Lange, R.,
    3. Vopper, G.,
    4. Cremer, H.,
    5. Heinlein, U. A. O.,
    6. Scheff, S.,
    7. Baldwin, S. A.,
    8. Leitges, M.,
    9. Cramer, M.,
    10. Paulsson, M. and
    11. et al
    . (1998). PACSIN, a brain protein that is upregulated upon differentiation into neuronal cells. Eur. J. Biochem 256, 201–211
    OpenUrlPubMedWeb of Science
    1. Qualmann, B.,
    2. Roos, J.,
    3. DiGregorio, P. J. and
    4. Kelly, R. B.
    (1999). Syndapin I, a synaptic dynamin-binding protein that associates with the neural Wiskott-Aldrich syndrome protein. Mol. Biol. Cell 10, 501–513
    OpenUrlAbstract/FREE Full Text
    1. Qualmann, B. and
    2. Kelly, R. B.
    (2000). Syndapin isoforms participate in receptor-mediated endocytosis and actin organisation. J. Cell Biol 148, 1047–1061
    OpenUrlAbstract/FREE Full Text
    1. Ringstad, N.,
    2. Nemoto, Y. and
    3. De Camilli, P.
    (1997). The SH3p4/SH3p8/SH3p13 protein family: binding partners for synaptojanin and dynamin via a Grb2-like Src homology 3 domain. Proc. Natl. Acad. Sci. USA 94, 8569–8574
    OpenUrlAbstract/FREE Full Text
    1. Ritter, B.,
    2. Modregger, J.,
    3. Paulsson, M. and
    4. Plomann, M.
    (1999). PACSIN 2, a novel member of the PACSIN family of cytoplasmic adapter proteins. FEBS Lett 454, 356–362
    OpenUrlCrossRefPubMedWeb of Science
    1. Schmid, S. L.,
    2. McNiven, M. A. and
    3. De Camilli, P.
    (1998). Dynamin and its partners: a progress report. Curr. Opin. Cell Biol 10, 504–512
    OpenUrlCrossRefPubMedWeb of Science
    1. Schmidt, A.,
    2. Wolde, M.,
    3. Thiele, C.,
    4. Fest, W.,
    5. Kratzin, H.,
    6. Podtelejnikov, A. V.,
    7. Witke, W.,
    8. Huttner, W. B. and
    9. Söling, H.-D.
    ) (1999). Endophilin I mediates synaptic vesicle formation by transfer of arachidonate to lysophophatic acid. Nature 401, 133–141
    OpenUrlCrossRefPubMedWeb of Science
    1. Sengar, A. S.,
    2. Wang, W.,
    3. Bishay, J.,
    4. Cohen, S. and
    5. Egan, S. E.
    (1999). The EH and SH3 domain Ese proteins regulate endocytosis by linking to dynamin and Eps15. EMBO J 18, 1159–1171
    OpenUrlAbstract
    1. Shupliakov, O.,
    2. Löw, P.,
    3. Grabs, D.,
    4. Gad, H.,
    5. Chen, H.,
    6. David, C.,
    7. Takei, K.,
    8. De Camilli, P. and
    9. Brodin, L.
    (1997). Synaptic vesicle endocytosis impaired by disruption of dynamin-SH3 domain interactions. Science 276, 259–263
    OpenUrlAbstract/FREE Full Text
    1. Sontag, J.-M.,
    2. Fykse, E. M.,
    3. Ushkaryov, Y.,
    4. Liu, J.-P.,
    5. Robinson, P. J. and
    6. Sudhof, T. C.
    (1994). Differential Expression and Regulation of Multiple Dynamins. J. Biol. Chem 269, 4547–4554
    OpenUrlAbstract/FREE Full Text
    1. Spencer, S.,
    2. Dowbenko, D.,
    3. Cheng, J.,
    4. Li, W.,
    5. Brush, J.,
    6. Utzig, S.,
    7. Simanis, V. and
    8. Lasky, L. A.
    (1997). PSTPIP: A tyrosine phosphorylated cleavage furrow-associated protein that is a substrate for a PEST tyrosine phosphatase. J. Cell Biol 138, 845–860
    OpenUrlAbstract/FREE Full Text
    1. Sudhof, T. C.,
    2. Czernik, A. J.,
    3. Kao, H. T.,
    4. Takei, K.,
    5. Johnston, P. A.,
    6. Horiuchi, A.,
    7. Kanazir, S. D.,
    8. Wagner, M. A.,
    9. Perin, M. S. and
    10. De Camilli, P.
    (1989). Synapsins: mosaics of shared and individual domains in a family of synaptic vesicle phosphoproteins. Science 245, 1474–1480
    OpenUrlAbstract/FREE Full Text
    1. Takei, Y.,
    2. Harada, A.,
    3. Takeda, S.,
    4. Kobayashi, K.,
    5. Terada, S.,
    6. Noda, T.,
    7. Takahashi, T. and
    8. Hirokawa, N.
    (1995). Synapsin I deficiency results in the structural change in the presynaptic terminals in the murine nervous system. J. Cell Biol 131, 1789–1800
    OpenUrlAbstract/FREE Full Text
    1. Urrutia, R.,
    2. Henley, J. R.,
    3. Cook, T. and
    4. McNiven, M. A.
    (1997). The dynamins: Redundant or distinct functions for an expanding family of related GTPases?. Proc. Natl. Acad. Sci. USA 94, 377–384
    OpenUrlAbstract/FREE Full Text
    1. Weng, Z.,
    2. Rickles, R. J.,
    3. Feng, S.,
    4. Richard, S.,
    5. Shaw, A. S.,
    6. Schreiber, S. L. and
    7. Brugge, J. S.
    (1995). Structure-function analysis of SH3 domains: SH3 binding specificity altered by single amino acid substitutions. Mol. Cell Biol 15, 5627–5634
    OpenUrlAbstract/FREE Full Text
    1. Wevers, A.,
    2. Burghaus, L.,
    3. Moser, N.,
    4. Witter, B.,
    5. Steinlein, O. K.,
    6. Schutz, U.,
    7. Achmitz, B.,
    8. Krempel, U.,
    9. Nowacki, S.,
    10. Pilz, K.,
    11. Stoodt, J.,
    12. Lindstrom, J.,
    13. De Vos, R. A.,
    14. Jansen Steur, E. N. and
    15. Schröder, H.
    (2000). Expression of nicotinic acetylcholine receptors in Alzheimer's disease: postmortem investigations and experimental approaches. Behav. Brain Res 113, 207–215
    OpenUrlCrossRefPubMedWeb of Science
    1. Wigge, P.,
    2. Vallis, Y. and
    3. McMahon, H. T.
    (1997). Inhibition of receptor-mediated endocytosis by the amphiphysin SH3 domain. Curr. Biol 7, 554–560
    OpenUrlCrossRefPubMedWeb of Science
    1. Wu, Y.,
    2. Spencer, S. D. and
    3. Lasky, L. A.
    (1998). Tyrosine phosphorylation regulates the SH3-mediated binding of the Wiskott-Aldrich syndrome protein to PSTPIP, a cytoskeletal-associated protein. J. Biol. Chem 273, 5765–5770
    OpenUrlAbstract/FREE Full Text
    1. Yamabhai, M.,
    2. Hoffman, N. G.,
    3. Hardison, N. L.,
    4. McPherson, P. S.,
    5. Castagnoli, L.,
    6. Cesareni, G. and
    7. Kay, B. K.
    (1998). Intersectin, a novel adaptor protein with two Eps15 homology and five Src homology 3 domains. J. Biol. Chem 273, 31401–31407
    OpenUrlAbstract/FREE Full Text
Back to top

 Download PDF

Email
Journal Article
All three PACSIN isoforms bind to endocytic proteins and inhibit endocytosis
J. Modregger, B. Ritter, B. Witter, M. Paulsson, M. Plomann
Journal of Cell Science 2000 113: 4511-4521;
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
Citation Tools
Alerts

Article navigation

Other journals from The Company of Biologists

Advertisement

Follow us on Instagram

Cell science is bursting with beautiful images and over on Instagram, we’re showing them off! Find both JCS and FocalPlane on Instagram for stories and techniques across cell biology.

An interview with Derek Walsh

Professor Derek Walsh is the guest editor of our new special issue Cell Biology of Host-Pathogen Interactions. In an interview, Derek tells us about his work in the field of DNA viruses, the impact of the pandemic on virology and what his role as Guest Editor taught him.

How to improve your scientific writing

"If you are a scientist and you want to succeed, you must become a writer."

How do scientists become master storytellers? We called on our journal Editors, proofreaders and contributors to our community sites for their advice on how to improve your scientific writing.

Meet the preLighters: Jennifer Ann Black

Following the theme of our latest special issue, postdoc Jennifer Ann Black studies replication stress and genome plasticity in Leishmania in Professor Luiz Tosi’s lab in Sao Paolo. We caught up with Jenn (virtually) to hear about her relocation to Brazil mid-pandemic, her research on parasites and what she enjoys about ‘preLighting’.

In our special issue, Chandrakar et al. and Rosazza et al. present their latest work on Leishmania.

Mole – The Corona Files

“There are millions of people around the world who continue to believe that the Terrible Pandemic is a hoax.”

Mole continues to offer his wise words to researchers on how to manage during the COVID-19 pandemic.

JCS and COVID-19

For more information on measures Journal of Cell Science is taking to support the community during the COVID-19 pandemic, please see here.

If you have any questions or concerns, please do not hestiate to contact the Editorial Office.