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Vol. 11, Issue 10, 3265-3275, October 2000
Department of Chemistry and Biochemistry, Howard Hughes Medical
Institute, University of Colorado, Boulder, Colorado 80309
Schizosaccharomyces pombe cells survive loss of
telomeres by a unique pathway of chromosome circularization. Factors
potentially involved in this survival mechanism include the
heterodimeric Ku protein and ligase IV, both of which are involved in
the repair of DNA double-strand breaks in mammalian cells. Furthermore,
Ku plays a role in telomere maintenance as well as in DNA double-strand break repair in Saccharomyces cerevisiae. We have
identified Ku and ligase IV homologues in S. pombe and
analyzed their functions during normal growth and in cells undergoing
senescence. In the absence of either a Ku subunit
(pku70+) or ligase IV
(lig4+), nonhomologous DNA end-joining was
severely reduced. Lack of functional Ku led to shorter but stable
telomeres and caused striking rearrangements of telomere-associated
sequences, indicating a function for Ku in inhibiting recombinational
activities near chromosome ends. In contrast to S.
cerevisiae, concurrent deletion of
pku70+ and the gene for the catalytic
subunit of telomerase (trt1+) was not
lethal, allowing for the first time the dissection of the roles of Ku
during senescence. Our results support a model in which Ku protects
chromosome termini from nucleolytic and recombinational activities but
is not involved in the formation of chromosome end fusions during
senescence. The conclusion that nonhomologous end-joining is not
required for chromosome circularization was further supported by
analysis of survivors in strains lacking the genes for both
trt1+ and lig4+.
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