Journal of Cell Science, Vol 112, Issue 10 1465-1476, Copyright © 1999 by Company of Biologists

JOURNAL ARTICLES

A novel hnRNP protein (HAP/SAF-B) enters a subset of hnRNP complexes and relocates in nuclear granules in response to heat shock

F Weighardt, F Cobianchi, L Cartegni, I Chiodi, A Villa, S Riva and G Biamonti
Istituto di Genetica Biochimica ed Evoluzionistica, CNR Via Abbiategrasso 207, Italy.

A two-hybrid screening in yeast for proteins interacting with the human hnRNP A1, yielded a nuclear protein of 917 amino acids that we termed hnRNP A1 associated protein (HAP). HAP contains an RNA binding domain (RBD) flanked by a negatively charged domain and by an S/K-R/E-rich region. In in vitro pull-down assays, HAP interacts with hnRNP A1, through its S/K-R/E-rich region, and with several other hnRNPs. HAP was found to be identical to the previously described Scaffold Attachment Factor B (SAF-B) and to HET, a transcriptional regulator of the Heat Shock Protein 27 gene. We show that HAP is a bona fide hnRNP protein, since anti-HAP antibodies immunoprecipitate from HeLa cell nucleoplasm the complete set of hnRNP proteins. Unlike most hnRNP proteins, the subnuclear distribution of HAP is profoundly modified in heat-shocked HeLa cells. Heat-shock treatment at 42 degrees C causes a transcription-dependent recruitment of HAP to a few large nuclear granules that exactly coincide with sites of accumulation of Heat Shock Factor 1 (HSF1). The recruitment of HAP to the granules is temporally delayed with respect to HSF1 and persists for a longer time during recovery at 37 degrees C. The hnRNP complexes immunoprecipitated from nucleoplasm of heat-shocked cells with anti-HAP antibodies have an altered protein composition with respect to canonical complexes. Altogether our results suggest an involvement of HAP in the cellular response to heat shock, possibly at the RNA metabolism level.

This article has been cited by other articles:

				R. Valgardsdottir, I. Chiodi, M. Giordano, A. Rossi, S. Bazzini, C. Ghigna, S. Riva, and G. Biamonti Transcription of Satellite III non-coding RNAs is a general stress response in human cells Nucleic Acids Res., February 2, 2008; 36(2): 423 - 434. [Abstract] [Full Text] [PDF]

				K. A. Sergeant, C. F. Bourgeois, C. Dalgliesh, J. P. Venables, J. Stevenin, and D. J. Elliott Alternative RNA splicing complexes containing the scaffold attachment factor SAFB2 J. Cell Sci., January 15, 2007; 120(2): 309 - 319. [Abstract] [Full Text] [PDF]

				C. Jolly and S. C. Lakhotia Human sat III and Drosophila hsr{omega} transcripts: a common paradigm for regulation of nuclear RNA processing in stressed cells Nucleic Acids Res., November 14, 2006; 34(19): 5508 - 5514. [Abstract] [Full Text] [PDF]

				K. M. Dobrzycka, K. Kang, S. Jiang, R. Meyer, P. H. Rao, A. V. Lee, and S. Oesterreich Disruption of Scaffold Attachment Factor B1 Leads to TBX2 Up-regulation, Lack of p19ARF Induction, Lack of Senescence, and Cell Immortalization Cancer Res., August 15, 2006; 66(16): 7859 - 7863. [Abstract] [Full Text] [PDF]

				Z Xiong, A Shaibani, Y-P Li, Y Yan, S Zhang, Y Yang, F Yang, H Wang, and X-F Yang Alternative splicing factor ASF/SF2 is down regulated in inflamed muscle J. Clin. Pathol., August 1, 2006; 59(8): 855 - 861. [Abstract] [Full Text] [PDF]

				M-B Debril, L Dubuquoy, J-N Feige, W Wahli, B Desvergne, J Auwerx, and L Gelman Scaffold attachment factor B1 directly interacts with nuclear receptors in living cells and represses transcriptional activity J. Mol. Endocrinol., December 1, 2005; 35(3): 503 - 517. [Abstract] [Full Text] [PDF]

				R. Valgardsdottir, I. Chiodi, M. Giordano, F. Cobianchi, S. Riva, and G. Biamonti Structural and Functional Characterization of Noncoding Repetitive RNAs Transcribed in Stressed Human Cells Mol. Biol. Cell, June 1, 2005; 16(6): 2597 - 2604. [Abstract] [Full Text] [PDF]

				M. Ivanova, K. M. Dobrzycka, S. Jiang, K. Michaelis, R. Meyer, K. Kang, B. Adkins, O. A. Barski, S. Zubairy, J. Divisova, et al. Scaffold Attachment Factor B1 Functions in Development, Growth, and Reproduction Mol. Cell. Biol., April 15, 2005; 25(8): 2995 - 3006. [Abstract] [Full Text] [PDF]

				K. Schmidt, J. Hoffend, A. Altmann, L. G. Strauss, A. Dimitrakopoulou-Strauss, B. Engelhardt, D. Koczan, J. Peter, S. Vorwald, H. Eskerski, et al. Transfer of the sFLT-1 Gene in Morris Hepatoma Results in Decreased Growth and Perfusion and Induction of Genes Associated with Stress Response Clin. Cancer Res., March 15, 2005; 11(6): 2132 - 2140. [Abstract] [Full Text] [PDF]

				A. Metz, J. Soret, C. Vourc'h, J. Tazi, and C. Jolly A key role for stress-induced satellite III transcripts in the relocalization of splicing factors into nuclear stress granules J. Cell Sci., September 1, 2004; 117(19): 4551 - 4558. [Abstract] [Full Text] [PDF]

				I. Chiodi, M. Corioni, M. Giordano, R. Valgardsdottir, C. Ghigna, F. Cobianchi, R.-M. Xu, S. Riva, and G. Biamonti RNA recognition motif 2 directs the recruitment of SF2/ASF to nuclear stress bodies Nucleic Acids Res., August 9, 2004; 32(14): 4127 - 4136. [Abstract] [Full Text] [PDF]

				S. M. Townson, K. Kang, A. V. Lee, and S. Oesterreich Structure-Function Analysis of the Estrogen Receptor {alpha} Corepressor Scaffold Attachment Factor-B1: IDENTIFICATION OF A POTENT TRANSCRIPTIONAL REPRESSION DOMAIN J. Biol. Chem., June 18, 2004; 279(25): 26074 - 26081. [Abstract] [Full Text] [PDF]

				P. G.N. Romano, P. Horton, and J. E. Gray The Arabidopsis Cyclophilin Gene Family Plant Physiology, April 1, 2004; 134(4): 1268 - 1282. [Abstract] [Full Text] [PDF]

				N. Rizzi, M. Denegri, I. Chiodi, M. Corioni, R. Valgardsdottir, F. Cobianchi, S. Riva, and G. Biamonti Transcriptional Activation of a Constitutive Heterochromatic Domain of the Human Genome in Response to Heat Shock Mol. Biol. Cell, February 1, 2004; 15(2): 543 - 551. [Abstract] [Full Text] [PDF]

				A. Sandqvist and L. Sistonen Nuclear stress granules: the awakening of a sleeping beauty? J. Cell Biol., January 5, 2004; 164(1): 15 - 17. [Abstract] [Full Text] [PDF]

				T.-P. Alastalo, M. Hellesuo, A. Sandqvist, V. Hietakangas, M. Kallio, and L. Sistonen Formation of nuclear stress granules involves HSF2 and coincides with the nucleolar localization of Hsp70 J. Cell Sci., September 1, 2003; 116(17): 3557 - 3570. [Abstract] [Full Text] [PDF]

				B. Sarkar, J.-Y. Lu, and R. J. Schneider Nuclear Import and Export Functions in the Different Isoforms of the AUF1/Heterogeneous Nuclear Ribonucleoprotein Protein Family J. Biol. Chem., May 30, 2003; 278(23): 20700 - 20707. [Abstract] [Full Text] [PDF]

				S. M. Townson, K. M. Dobrzycka, A. V. Lee, M. Air, W. Deng, K. Kang, S. Jiang, N. Kioka, K. Michaelis, and S. Oesterreich SAFB2, a New Scaffold Attachment Factor Homolog and Estrogen Receptor Corepressor J. Biol. Chem., May 23, 2003; 278(22): 20059 - 20068. [Abstract] [Full Text] [PDF]

				A. Traweger, R. Fuchs, I. A. Krizbai, T. M. Weiger, H.-C. Bauer, and H. Bauer The Tight Junction Protein ZO-2 Localizes to the Nucleus and Interacts with the Heterogeneous Nuclear Ribonucleoprotein Scaffold Attachment Factor-B J. Biol. Chem., January 17, 2003; 278(4): 2692 - 2700. [Abstract] [Full Text] [PDF]

				M. Denegri, D. Moralli, M. Rocchi, M. Biggiogera, E. Raimondi, F. Cobianchi, L. De Carli, S. Riva, and G. Biamonti Human Chromosomes 9, 12, and 15 Contain the Nucleation Sites of Stress-Induced Nuclear Bodies Mol. Biol. Cell, June 1, 2002; 13(6): 2069 - 2079. [Abstract] [Full Text] [PDF]

				C. Jolly, L. Konecny, D. L. Grady, Y. A. Kutskova, J. J. Cotto, R. I. Morimoto, and C. Vourc'h In vivo binding of active heat shock transcription factor 1 to human chromosome 9 heterochromatin during stress J. Cell Biol., March 4, 2002; 156(5): 775 - 781. [Abstract] [Full Text] [PDF]

				M. Denegri, I. Chiodi, M. Corioni, F. Cobianchi, S. Riva, and G. Biamonti Stress-induced Nuclear Bodies Are Sites of Accumulation of Pre-mRNA Processing Factors Mol. Biol. Cell, November 1, 2001; 12(11): 3502 - 3514. [Abstract] [Full Text] [PDF]

				E. Nikolakaki, R. Kohen, A. M. Hartmann, S. Stamm, E. Georgatsou, and T. Giannakouros Cloning and Characterization of an Alternatively Spliced Form of SR Protein Kinase 1 That Interacts Specifically with Scaffold Attachment Factor-B J. Biol. Chem., October 19, 2001; 276(43): 40175 - 40182. [Abstract] [Full Text] [PDF]

				O. Nayler, A. M. Hartmann, and S. Stamm The ER Repeat Protein YT521-B Localizes to a Novel Subnuclear Compartment J. Cell Biol., September 5, 2000; 150(5): 949 - 962. [Abstract] [Full Text] [PDF]

				I.-E. Gallouzi, C. M. Brennan, M. G. Stenberg, M. S. Swanson, A. Eversole, N. Maizels, and J. A. Steitz HuR binding to cytoplasmic mRNA is perturbed by heat shock PNAS, March 28, 2000; 97(7): 3073 - 3078. [Abstract] [Full Text] [PDF]

				I Chiodi, M Biggiogera, M Denegri, M Corioni, F Weighardt, F Cobianchi, S Riva, and G Biamonti Structure and dynamics of hnRNP-labelled nuclear bodies induced by stress treatments J. Cell Sci., January 11, 2000; 113(22): 4043 - 4053. [Abstract] [PDF]

				O. Stoss, M. Olbrich, A. M. Hartmann, H. Konig, J. Memmott, A. Andreadis, and S. Stamm The STAR/GSG Family Protein rSLM-2 Regulates the Selection of Alternative Splice Sites J. Biol. Chem., March 16, 2001; 276(12): 8665 - 8673. [Abstract] [Full Text] [PDF]

				C. Jolly, L. Konecny, D. L. Grady, Y. A. Kutskova, J. J. Cotto, R. I. Morimoto, and C. Vourc'h In vivo binding of active heat shock transcription factor 1 to human chromosome 9 heterochromatin during stress J. Cell Biol., March 4, 2002; 156(5): 775 - 781. [Abstract] [Full Text] [PDF]