Cytostellin distributes to nuclear regions enriched with splicing factors

生物 剪接体 RNA剪接 前体mRNA 相间 细胞生物学 细胞周期 有丝分裂 细胞核 SR蛋白 核蛋白 snRNP公司 核糖核酸 分子生物学 细胞 细胞质 遗传学 转录因子 基因
作者
David B. Bregman,Lei Du,Li Yi,Stephen Ribisi,Stephen L. Warren
出处
期刊:Journal of Cell Science [The Company of Biologists]
卷期号:107 (3): 387-396 被引量:34
标识
DOI:10.1242/jcs.107.3.387
摘要

ABSTRACT Cytostellin, a ∼240 kDa phosphoprotein found in all cells examined from human to yeast, is predominantly intranuclear in interphase mammalian cells and undergoes continuous redistribution during the cell cycle. Here, mammalian cytostellin is shown to localize to intranuclear regions enriched with multiple splicing proteins, including spliceosome assembly factor, SC-35. Cytostellin and the splicing proteins also co-localize to discrete foci (called ‘dots’), which are distributed throughout the cell during mitosis and part of G1. The cytostellin that is localized to these dots resists extraction by Triton X-100, indicating that it is tightly associated with insoluble cell structures. All immunostainable cytostellin reappears in the nucleus before S-phase. Although cytostellin and the splicing proteins co-localize in interphase and dividing cells, cytostellin is not detected in purified spliceosomes, and it associates with six unidentified proteins, forming a macro-molecular complex that is biochemically distinct from the proteins that comprise spliceosomes. This macromolecular complex is detected at constant levels throughout the cell cycle, and the level of cytostellin protein remains constant during the cell cycle. Nevertheless, intranuclear cytostellin immunostaining fluctuates markedly during the cell cycle. The monoclonal antibody (mAb) H5 epitope of cytostellin is ‘masked’ in serum-starved cells, but 60 minutes after serum stimulation intense cytostellin immunoreactivity appears in the nuclear speckles. This rapid induction of cytostellin immunoreactivity in subnuclear regions enriched with many splicing factors, as well as accumulations of RNA polymerase II (Pol II) transcripts, suggests that cytostellin may have a function related to mRNA biogenesis.

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