Functional association of human Ki-1/57 with pre-mRNA splicing events

被引:15
作者
Bressan, Gustavo C. [2 ]
Quaresma, Alexandre J. C.
Moraes, Eduardo C. [2 ]
Manfiolli, Adriana O. [3 ]
Passos, Dario O.
Gomes, Marcelo D. [3 ]
Kobarg, Joerg [1 ,2 ]
机构
[1] Ctr Biol Mol Estrutural, Lab Nacl Luz Sincrotron, BR-13084971 Campinas, SP, Brazil
[2] Univ Estadual Campinas, Inst Biol, Campinas, SP, Brazil
[3] Univ Sao Paulo, Fac Med Ribeirao Preto, Dept Bioquim & Imunol, Ribeirao Preto, Brazil
基金
巴西圣保罗研究基金会;
关键词
Arg methylation; nuclear bodies; protein-protein interaction; RNA binding; splicing; REGULATORY PROTEIN KI-1/57; MUSCULAR-ATROPHY PROTEIN; C-TERMINAL REGION; BINDING-PROTEIN; ARGININE METHYLTRANSFERASES; COILED BODIES; SMN COMPLEX; METHYLATION; INTERACTS; NUCLEUS;
D O I
10.1111/j.1742-4658.2009.07092.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
070307 [化学生物学]; 071010 [生物化学与分子生物学];
摘要
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 Ki- 1 / 57 with RNA functions. Through immunoprecipitation assays, we verified the association of Ki- 1 / 57 with the endogenous splicing proteins hnRNPQ and SFRS9 in HeLa cell extracts. We also found that recombinant Ki- 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 Ki- 1 / 57 can modify the splicing site selection of the adenoviral E1A minigene in a dose- dependent manner. Further confocal and. uorescence microscopy analysis revealed the localization of enhanced green. uorescent 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, our findings suggest that Ki- 1 / 57 is probably involved in cellular events related to RNA functions, such as pre- mRNA splicing.
引用
收藏
页码:4194 / 4207
页数:14
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