Genetics, Vol. 150, 1019-1035, November 1998, Copyright © 1998

Accumulation of mRNA Coding for the Ctf13p Kinetochore Subunit of Saccharomyces cerevisiae Depends on the Same Factors That Promote Rapid Decay of Nonsense mRNAs

Jeffrey N. Dahlseida, John Puzissb, Renee L. Shirleya, Audrey L. Atkinc, Philip Hieterd, and Michael R. Culbertsona
a Laboratories of Genetics and Molecular Biology, University of Wisconsin, Madison, Wisconsin 53706,
b Department of Microbiology, Bristol-Myers Squibb Pharmaceutical Research Institute, Wallingford, Connecticut 06492,
c School of Biological Sciences, University of Nebraska, Lincoln, Nebraska 68588
d Centre for Molecular Medicine and Therapeutics, University of British Columbia, Vancouver, British Columbia V5Z 4H4 Canada

Corresponding author: Michael R. Culbertson, Laboratory of Molecular Biology, University of Wisconsin-Madison, 413 R.M. Bock Laboratories, 1525 Linden Dr., Madison, WI 53706-1596., mrculber{at}facstaff.wisc.edu (E-mail).

Communicating editor: E. W. JONES

The CTF13 gene codes for a subunit of the kinetochore in Saccharomyces cerevisiae. The temperature-sensitive mutation ctf13-30, which confers reduced fidelity of chromosome transmission, is a G -> A transition causing an amino acid substitution of Lys for Glu146. Strains carrying one chromosomal copy of ctf13-30 fail to grow at the restrictive temperature, whereas a haploid strain carrying two copies of ctf13-30 can grow. Four genes, UPF1, UPF2, UPF3, and ICK1, were represented among extragenic suppressors of ctf13-30. The UPF genes encode proteins that promote rapid decay of pre-mRNAs and mRNAs containing a premature stop codon. Suppressor mutations in these genes restore kinetochore function by causing increased accumulation of ctf13-30 mRNA. They also cause increased accumulation of CYH2 pre-mRNA, which is a natural target of UPF-mediated decay. Mutations in ICK1 restore kinetochore function but have no effect on ctf13-30 mRNA or CYH2 pre-mRNA accumulation. Most importantly, loss of UPF1 function causes increased accumulation of wild-type CTF13 mRNA but has no effect on the mRNA half-life. We propose that UPF-mediated decay modulates the mRNA level of one or more factors involved in CTF13 mRNA expression.




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