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Published online 3 November 2003. doi:10.1083/jcb.200306132
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© The Rockefeller University Press, 0021-9525/2003/11/457 $8.00
The Journal of Cell Biology, Volume 163, Number 3, 457-461

Report

A function for the mitochondrial chaperonin Hsp60 in the structure and transmission of mitochondrial DNA nucleoids in Saccharomyces cerevisiae



Brett A. Kaufman, Jill E. Kolesar, Philip S. Perlman and Ronald A. Butow

Department of Molecular Biology, The University of Texas Southwestern Medical Center, Dallas, TX 75390

Address correspondence to Ronald Butow, Department of Molecular Biology, The University of Texas Southwestern Medical Center, 5323 Harry Hines Blvd., Dallas, TX 75390-9148. Tel.: (214) 648-1465. Fax: (214) 648-1488. email: ronald.butow{at}utsouthwestern.edu

The yeast mitochondrial chaperonin Hsp60 has previously been implicated in mitochondrial DNA (mtDNA) transactions: it is found in mtDNA nucleoids associated with single-stranded DNA; it binds preferentially to the template strand of active mtDNA ori sequences in vitro; and wild-type ({rho}+) mtDNA is unstable in hsp60 temperature-sensitive (ts) mutants grown at the permissive temperature. Here we show that the mtDNA instability is caused by a defect in mtDNA transmission to daughter cells. Using high resolution, fluorescence deconvolution microscopy, we observe a striking alteration in the morphology of mtDNA nucleoids in {rho}+ cells of an hsp60-ts mutant that suggests a defect in nucleoid division. We show that {rho}- petite mtDNA consisting of active ori repeats is uniquely unstable in the hsp60-ts mutant. This instability of ori {rho}- mtDNA requires transcription from the canonical promoter within the ori element. Our data suggest that the nucleoid dynamics underlying mtDNA transmission are regulated by the interaction between Hsp60 and mtDNA ori sequences.

Key Words: yeast; mitochondria; mitochondrial DNA, nucleoids; Hsp60

B.A. Kaufman's present address is Montreal Neurological Institute, 3801 University St., Montreal, Quebec H3A 2B4, Canada.

Abbreviations used in this paper: mtDNA, mitochondrial DNA; ss, single stranded; ts, temperature sensitive.


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