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Molecular and Cellular Biology, March 2000, p. 1816-1824, Vol. 20, No. 5
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

A DNA Helicase Required for Maintenance of the Functional Mitochondrial Genome in Saccharomyces cerevisiae

Tiina Sedman,1 Silja Kuusk,1 Sirje Kivi,1 and Juhan Sedman2,*

Institute of Molecular and Cellular Biology, Tartu University,1 and Estonian Biocentre,2 Tartu 51010, Estonia

Received 21 October 1999/Accepted 24 November 1999

A novel DNA helicase, a homolog of several prokaryotic helicases, including Escherichia coli Rep and UvrD proteins, is encoded by the Saccharomyces cerevisiae nuclear genome open reading frame YOL095c on the chromosome XV. Our data demonstrate that the helicase is localized in the yeast mitochondria and is loosely associated with the mitochondrial inner membrane during biochemical fractionation. The sequence of the C-terminal end of the 80-kDa helicase protein is similar to a typical N-terminal mitochondrial targeting signal; deletions and point mutations in this region abolish transport of the protein into mitochondria. The C-terminal signal sequence of the helicase targets a heterologous carrier protein into mitochondria in vivo. The purified recombinant protein can unwind duplex DNA molecules in an ATP-dependent manner. The helicase is required for the maintenance of the functional ([rho+]) mitochondrial genome on both fermentable and nonfermentable carbon sources. However, the helicase is not essential for the maintenance of several defective ([rho-]) mitochondrial genomes. We also demonstrate that the helicase is not required for transcription in mitochondria.

* Corresponding author. Mailing address: Estonian Biocentre, Riia 23, Tartu 51010, Estonia. Phone: 372-7-375037. Fax: 372-7-420286. E-mail: jsedman{at}ebc.ee.

Molecular and Cellular Biology, March 2000, p. 1816-1824, Vol. 20, No. 5
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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