Genetics, Vol 131, 255-260, Copyright © 1992

INVESTIGATIONS

Nuclear Mutations in the Petite-Negative Yeast Schizosaccharomyces pombe Allow Growth of Cells Lacking Mitochondrial DNA

P. Haffter and T. D. Fox
Present address: Max-Planck-Institut fur Entwicklungsbiologie, Spemannstrasse 35/III, 7400 Tubingen, Germany.

The fission yeast Schizosaccharomyces pombe has never been found to give rise to viable cells totally lacking mitochondrial DNA (rho{deg}). This paper describes the isolation of rho{deg} strains of S. pombe by very long term incubation of cells in liquid medium containing glucose, potassium acetate and ethidium bromide. Once isolated, the rho{deg} strains did not require potassium acetate or any other novel growth factors. These nonrespiring strains contained no mitochondrial DNA (mtDNA) detectable either by gel-blot hybridization using as probe a clone containing the entire S. pombe mtDNA, or by 1',6-diamidino-2-phenylindole staining of whole cells. Induction of rho{deg} derivatives of standard laboratory strains was not reproducible from culture to culture. The cause of this irreproducibility appears to be that growth of the rho{deg} strains of S. pombe depended on nuclear mutations that occurred in some, but not all, of the initial cultures. Two independent rho{deg} isolates contained mutations in unlinked genes, termed ptp1-1 and ptp2-1. These mutations allowed reproducible ethidium bromide induction of viable rho{deg} strains. No other phenotypes were associated with ptp mutations in rho(+) strains.

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