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A more recent version of this article appeared on January 1, 2004
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Submitted on July 20, 2003
Revised on September 8, 2003
Accepted on September 24, 2003
1 Cellular & Molecular Biology Laboratory, RIKEN, Hirosawa 2-1, Wako-shi, Saitama 351-0198, Japan, and CREST, Japan Science and Technology Corporation (JST)
* Corresponding author. E-mail address: tshibata{at}postman.riken.go.jp.
Mitochondria carry many copies of mitochondrial (mt) DNA, but mt-alleles quickly segregate during mitotic growth through unknown mechanisms. Consequently, all mtDNA copies are often genetically homogeneous within each individual ("homoplasmic"). Our previous study suggested that tandem multimers ("concatemers") formed mainly by the Mhr1p (a yeast nuclear gene encoded mtDNA-recombination protein)-dependent pathway are required for mtDNA-partitioning into buds with concomitant monomerization. The transmission of a few randomly selected clones (as concatemers) of mtDNA into buds is a possible mechanism to establish homoplasmy. The current study provides evidence for this hypothesis as follows: the overexpression of MHR1 accelerates mt-allele-segregation in growing heteroplasmic zygotes, and mhr1-1 (recombination-deficient) causes its delay. The mt-allele-segregation rate correlates with the abundance of concatemers, which depends on Mhr1p. In G1-arrested cells, concatemeric mtDNA was labeled by [14C]thymidine at a much higher density than monomers, indicating concatemers as the immediate products of mtDNA-replication, most likely in a rolling-circle mode. After releasing the G1-arrest in the absence of [14C]thymidine, the monomers as the major species in growing buds of dividing cells bear a similar density of 14C as the concatemers in the mother cells, indicating that the concatemers in mother cells are the precursors of the monomers in buds.
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