Department of Molecular Biotechnology, Graduate School of Advanced Sciences of Matter, Hiroshima University, 1-3-1 Kagamiyama, Higashi-Hiroshima 739-8530, Japan.
Mol Cell Biol. 2011 Feb;31(3):495-506. doi: 10.1128/MCB.00613-10. Epub 2010 Nov 22.
Pot1 is a single-stranded telomere-binding protein that is conserved from fission yeast to mammals. Deletion of Schizosaccharomyces pombe pot1(+) causes immediate telomere loss. S. pombe Rqh1 is a homolog of the human RecQ helicase WRN, which plays essential roles in the maintenance of genomic stability. Here, we demonstrate that a pot1Δ rqh1-hd (helicase-dead) double mutant maintains telomeres that are dependent on Rad51-mediated homologous recombination. Interestingly, the pot1Δ rqh1-hd double mutant displays a "cut" (cell untimely torn) phenotype and is sensitive to the antimicrotubule drug thiabendazole (TBZ). Moreover, the chromosome ends of the double mutant do not enter the pulsed-field electrophoresis gel. These results suggest that the entangled chromosome ends in the pot1Δ rqh1-hd double mutant inhibit chromosome segregation, signifying that Pot1 and Rqh1 are required for efficient chromosome segregation. We also found that POT1 knockdown, WRN-deficient human cells are sensitive to the antimicrotubule drug vinblastine, implying that some of the functions of S. pombe Pot1 and Rqh1 may be conserved in their respective human counterparts POT1 and WRN.
Pot1 是一种单链端粒结合蛋白,从裂殖酵母到哺乳动物都有保守性。Schizosaccharomyces pombe pot1(+)的缺失会导致端粒立即丢失。S. pombe Rqh1 是人类 RecQ 解旋酶 WRN 的同源物,在维持基因组稳定性方面发挥着重要作用。在这里,我们证明了 pot1Δ rqh1-hd(解旋酶失活)双突变体维持依赖 Rad51 介导的同源重组的端粒。有趣的是,pot1Δ rqh1-hd 双突变体表现出“切割”(细胞过早撕裂)表型,并且对微管药物噻苯达唑(TBZ)敏感。此外,双突变体的染色体末端不进入脉冲场电泳凝胶。这些结果表明,pot1Δ rqh1-hd 双突变体中缠绕的染色体末端抑制染色体分离,表明 Pot1 和 Rqh1 是有效染色体分离所必需的。我们还发现,POT1 敲低,WRN 缺陷的人类细胞对微管药物长春碱敏感,这意味着裂殖酵母 Pot1 和 Rqh1 的某些功能可能在其相应的人类对应物 POT1 和 WRN 中保守。