Huang Han-Kuei, Bailis Julie M, Leverson Joel D, Gómez Eliana B, Forsburg Susan L, Hunter Tony
Molecular and Cell Biology Laboratory, The Salk Institute for Biological Studies, La Jolla, CA 92037, USA.
Mol Cell Biol. 2005 Oct;25(20):9000-15. doi: 10.1128/MCB.25.20.9000-9015.2005.
Fission yeast Bir1p/Cut17p/Pbh1p, the homolog of human Survivin, is a conserved chromosomal passenger protein that is required for cell division and cytokinesis. To study how Bir1p promotes accurate segregation of chromosomes, we generated and analyzed a temperature-sensitive allele, bir1-46, and carried out genetic screens to find genes that interact with bir1(+). We identified Psf2p, a component of the GINS complex required for DNA replication initiation, as a high-copy-number suppressor of the bir1-46 growth defect. Loss of Psf2p function by depletion or deletion or by use of a temperature-sensitive allele, psf2-209, resulted in chromosome missegregation that was associated with mislocalization of Bir1p. We also found that the human homolog of Psf2p, PSF2, was required for proper chromosome segregation. In addition, we observed that high-copy-number expression of Pic1p, the fission yeast homolog of INCENP (inner centromere protein), suppressed bir1-46. Pic1p exhibited a localization pattern typical of chromosomal passenger proteins. Deletion of pic1(+) caused chromosome missegregation phenotypes similar to those of bir1-46. Our data suggest that Bir1p and Pic1p act as part of a conserved chromosomal passenger complex and that Psf2p/GINS indirectly affects the localization and function of this complex in chromosome segregation, perhaps through an S-phase role in centromere replication.
裂殖酵母中的Bir1p/Cut17p/Pbh1p是人类Survivin的同源物,是一种保守的染色体乘客蛋白,对于细胞分裂和胞质分裂是必需的。为了研究Bir1p如何促进染色体的准确分离,我们构建并分析了一个温度敏感等位基因bir1-46,并进行了遗传筛选以寻找与bir1(+)相互作用的基因。我们鉴定出Psf2p,一种DNA复制起始所需的GINS复合物的组成成分,作为bir1-46生长缺陷的高拷贝数抑制因子。通过缺失或耗尽Psf2p功能,或使用温度敏感等位基因psf2-209,会导致染色体错分离,这与Bir1p的定位错误相关。我们还发现Psf2p的人类同源物PSF2对于正确的染色体分离是必需的。此外,我们观察到裂殖酵母中INCENP(着丝粒内蛋白)的同源物Pic1p的高拷贝数表达抑制了bir1-46。Pic1p表现出典型的染色体乘客蛋白的定位模式。缺失pic1(+)会导致与bir1-46相似的染色体错分离表型。我们的数据表明,Bir1p和Pic1p作为保守的染色体乘客复合物的一部分发挥作用,并且Psf2p/GINS可能通过在着丝粒复制中的S期作用间接影响该复合物在染色体分离中的定位和功能。