Nucleic Acids Research, Vol 24, Issue 23 4676-4683, Copyright © 1996 by Oxford University Press
JP Javerzat, G Cranston and RC Allshire
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.
ARTICLES
Fission yeast genes which disrupt mitotic chromosome segregation when overexpressed
M.R.C. Human Genetics Unit, Western General Hospital, Edinburgh, UK. jpaul.javerzat@ibgc.u-bordeaux2.fr
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