Fission yeast genes which disrupt mitotic chromosome segregation when overexpressed

被引:57
作者
Javerzat, JP [1 ]
Cranston, G [1 ]
Allshire, RC [1 ]
机构
[1] WESTERN GEN HOSP, HUMAN GENET UNIT, MRC, EDINBURGH EH4 2XU, MIDLOTHIAN, SCOTLAND
基金
英国惠康基金;
关键词
D O I
10.1093/nar/24.23.4676
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
An interference assay has been devised in Schizosaccharomyces pombe to rapidly identify and clone genes involved in chromosome segregation, Random S.pombe cDNAs were overexpressed from an inducible promoter in a strain carrying an additional, non-essential minichromosome, Overexpression of cDNAs derived from four genes, two known (nda3(+) and ubc4(+), encoding beta-tubulin and a ubiquitin conjugating enzyme, respectively) and two unknown, named mlo2(+) and mlo3(+) (missegregation & lethal when over expressed) caused phenotypes consistent with a failure to segregate chromosomes, Full overexpression of all four cDNAs was lethal, Cells overexpressing nda3(+) and ubc4(+) cDNAs arrested with condensed unsegregated chromosomes and cells overexpressing mlo2(+) displayed an asymmetric distribution of nuclear chromatin, Sublethal levels of overexpression of nda3(+), ubc4(+) and mlo2(+) cDNAs caused elevated rates of minichromosome loss, A third cDNA mlo3(+), displayed no increase in the frequency of minichromosome loss at sublethal levels of overexpression but full overexpression caused a complete failure to segregate chromosomes, Our results confirm the assumption that beta-tubulin overexpression is lethal in S.pombe, implicate ubc4(+) in the control of metaphase-anaphase transition in fission yeast and finally identify two new genes, mlo2(+) and mlo3(+), likely to play an important role for chromosome transmission fidelity in mitosis.
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页码:4676 / 4683
页数:8
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