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© The Rockefeller University Press, 0021-9525/1999//15 $5.00
The Journal of Cell Biology, Volume 145, Number 1, , 1999 15-28

Regular Articles

Ctf19p: A Novel Kinetochore Protein in Saccharomyces cerevisiae and a Potential Link between the Kinetochore and Mitotic Spindle



Katherine M. Hyland*,{ddagger}, Jeffrey Kingsbury§, Doug Koshland§, and Philip Hieter{ddagger}

* The Johns Hopkins University School of Medicine, Department of Molecular Biology and Genetics, Baltimore, Maryland 21205; {ddagger} Centre for Molecular Medicine and Therapeutics, University of British Columbia, Vancouver, British Columbia V5Z 4H4, Canada; and § Carnegie Institute of Washington, Department of Embryology, Baltimore, Maryland 21210

A genetic synthetic dosage lethality (SDL) screen using CTF13 encoding a known kinetochore protein as the overexpressed reference gene identified two chromosome transmission fidelity (ctf) mutants, YCTF58 and YCTF26. These mutant strains carry independent alleles of a novel gene, which we have designated CTF19. In light of its potential role in kinetochore function, we have cloned and characterized the CTF19 gene in detail. CTF19 encodes a nonessential 369–amino acid protein. ctf19 mutant strains display a severe chromosome missegregation phenotype, are hypersensitive to benomyl, and accumulate at G2/M in cycling cells. CTF19 genetically interacts with kinetochore structural mutants and mitotic checkpoint mutants. In addition, ctf19 mutants show a defect in the ability of centromeres on minichromosomes to bind microtubules in an in vitro assay. In vivo cross-linking and chromatin immunoprecipitation demonstrates that Ctf19p specifically interacts with CEN DNA. Furthermore, Ctf19-HAp localizes to the nuclear face of the spindle pole body and genetically interacts with a spindle-associated protein. We propose that Ctf19p is part of a macromolecular kinetochore complex, which may func- tion as a link between the kinetochore and the mitotic spindle.

Key Words: centromere • kinetochores • chromosome segregation • mitotic spindle apparatus • Saccharomyces cerevisiae

Abbreviations used in this paper: budding uninhibited by benzimidazole; CDEs, centromere DNA elements; ctf, chromosome transmission fidelity; 5-FOA, 5-fluoroorotic acid; mad, mitotic arrest deficient; MT, microtubule; NZ, nocodazole; ORF, open reading frame; SDL, synthetic dosage lethality; SL, synthetic lethality; SPB, spindle pole body.


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