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The cytoplasmic and nuclear protein Ki-1⁄57 was first identified in malig-nant cells from Hodgkin’s lymphoma. Despite studies showing its phos-phorylation, arginine methylation, and interaction with several regulatory proteins, the functional role of Ki-1⁄57 in human cells remains to be determined. | Functional association of human Ki-1 57 with pre-mRNA splicing events Gustavo C. Bressan1 2 Alexandre J. C. Quaresma1 Eduardo C. Moraes1 2 Adriana O. Manfiolli3 Dario O. Passos1 Marcelo D. Gomes3 and Jorg Kobarg1 2 1 Laboratorio Nacionalde Luz Sincrotron Campinas SP Brasil 2 Instituto de Biologia Universidade Estadualde Campinas Campinas SP Brasil 3 Departamento de Bioquimica e Imunologia Faculdade de Medicina de Ribeirao Preto da Universidade de Scio Paulo Ribeirao Preto Brasil Keywords Arg methylation nuclear bodies protein-protein interaction RNA binding splicing Correspondence J. Kobarg Laboratorio Nacionalde Luz Sincrotron Centro de Biologia Molecular Estrutural Rua Giuseppe Maximo Scolfaro 10.000 C.P. 6192 13084-971 Campinas SP Brasil Fax 55 19 3512 1006 Tel 55 19 3512 1125 E-mail jkobarg@lnls.br Received 12 April2009 revised 8 May 2009 accepted 13 May 2009 doi 10.1111 j.1742-4658.2009.07092.x The cytoplasmic and nuclear protein Ki-1 57 was first identified in malignant cells from Hodgkin s lymphoma. Despite studies showing its phosphorylation arginine methylation and interaction with several regulatory proteins the functional role of Ki-1 57 in human cells remains to be determined. Here we investigated the relationship of KÍ-1 57 with RNA functions. Through immunoprecipitation assays we verified the association of KÍ-1 57 with the endogenous splicing proteins hnRNPQ and SFRS9 in HeLa cell extracts. We also found that recombinant KÍ-1 57 was able to bind to a poly-U RNA probe in electrophoretic mobility shift assays. In a classic splicing test we showed that KÍ-1 57 can modify the splicing site selection of the adenoviral E1A minigene in a dose-dependent manner. Further confocal and fluorescence microscopy analysis revealed the localization of enhanced green fluorescent protein-Ki-1 57 to nuclear bodies involved in RNA processing and or small nuclear ribonucleoprotein assembly depending on the cellular methylation status and its N-terminal region. In summary
