Functional and structural studies of three human regulatory proteins: Fez1, Ki-1/57 and NSAP1 (hnRNP-Q)

• Grant number: 05/00235-1
• Support Opportunities: Regular Research Grants
• Start date: August 01, 2005
• End date: October 31, 2007
• Field of knowledge: Biological Sciences - Biochemistry
• Principal Investigator: Jörg Kobarg
• Grantee: Jörg Kobarg
• Host Institution: Associação Brasileira de Tecnologia de Luz Síncrotron (ABTLuS). Ministério da Ciência, Tecnologia e Inovação (Brasil). Campinas , SP, Brazil

Abstract

We want to study the function and structure of three human regulatory proteins which are involved in human diseases. About the protein NSAP1 (hnRNP-Q) it is known that it is a RNA binding protein and that it is possibly involved in pre-mRNA - splicing - and the degradation of mRNAs, which encode oncoproteins. Structurally this protein was not studied yet, but it is known that it is composed of one - acid domain, three RRM domains (RNA recognition motifs, which bind RNA) and one RG-box, which a target of arginine methylation. The functions and structures of the other two proteins, Ki-1/57 and Fez1, are still not known. In previous studies we identified a series of proteins that are involved in cellular signalization processes and the regulation of transcription, that interact with Fez1 or Ki-1/57. This suggests that both proteins are involved in this cellular context, although the association to other cellular process cannot be ruled out. In this project we intend to study the structure of the mentioned proteins or of their individual domains by spectroscopic methods and protein cristallography. We also will study the function of these proteins using several modern molecular biology techniques, including among others: gene silencing (iRNA method), identification of post-translational modifications and interacting proteins in vitro (mass spectrometry) as well as in vitro (yeast two-hybrid system), SELEX (Systematic evolution of ligands by exponential enrichment) to identify interacting RNA target sequences. (AU)

Scientific publications

(5)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

LANZA‚ D.C.F.; TRINDADE‚ D.M.; ASSMANN‚ E.M.; KOBARG‚ J.. Over-expression of GFP-FEZ1 causes generation of multi-lobulated nuclei mediated by microtubules in HEK293 cells. Experimental Cell Research, v. 314, n. 10, p. 2028-2039, 2008. (05/00235-1)

ASSMANN, ELIANA M.; ALBORGHETTI, MARCOS R.; CAMARGO, MARIA E. R.; KOBARG, JORG. FEZ1 dimerization and interaction with transcription regulatory proteins involves its coiled-coil region. Journal of Biological Chemistry, v. 281, n. 15, p. 9869-9881, Apr. 2006. (05/00235-1)

LANZA, DANIEL C. F.; SILVA, JULIO C.; ASSMANN, ELIANA M.; QUARESMA, ALEXANDRE J. C.; BRESSAN, GUSTAVO C.; TORRIANI, IRIS L.; KOBARG, JOERG. Human FEZ1 has characteristics of a natively unfolded protein and dimerizes in solution. PROTEINS-STRUCTURE FUNCTION AND BIOINFORMATICS, v. 74, n. 1, p. 104-121, JAN 2009. (05/00235-1)

QUARESMA, ALEXANDRE J. C.; BRESSAN, G. C.; GAVA, L. M.; LANZA, D. C. F.; RAMOS, C. H. I.; KOBARG, JOERG. Human hnRNP Q re-localizes to cytoplasmic granules upon PMA, thapsigargin, arsenite and heat-shock treatments. Experimental Cell Research, v. 315, n. 6, p. 968-980, APR 1 2009. (05/00235-1)

BRESSAN, GUSTAVO C.; SILVA, JULIO C.; BORGES, JULIO C.; DOS PASSOS, DARIO O.; RAMOS, CARLOS H. I.; TORRIANI, IRIS L.; KOBARG, JOERG. Human regulatory protein Ki-1/57 has characteristics of an intrinsically unstructured protein. JOURNAL OF PROTEOME RESEARCH, v. 7, n. 10, p. 4465-4474, OCT 2008. (05/00235-1)