Unlinked Noncomplementation: Isolation of New Conditional-Lethal Mutations in Each of the Tubulin Genes of Saccharomyces cerevisiae

INVESTIGATIONS

Unlinked Noncomplementation: Isolation of New Conditional-Lethal Mutations in Each of the Tubulin Genes of Saccharomyces cerevisiae

T. Stearns and D. Botstein
Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139

Mutations in genes of Saccharomyces cerevisiae that code for proteins that interact with {beta}-tubulin were sought by screening for unlinked mutations that fail to complement mutations in the single {beta}-tubulin-encoding gene (TUB2). Among the first three noncomplementing mutations examined, two are linked to TUB2 while one is unlinked. The unlinked mutation was shown to be a conditional-lethal allele of the major {alpha}-tubulin-encoding gene (TUB1) and represents the first such mutation in that gene. The tub1-1 mutation itself causes a cold-sensitive cell-cycle arrest, and confers supersensitivity to the antimicrotubule drug benomyl. These phenotypes occur in the presence of a wild-type copy of the minor {alpha}-tubulin-encoding gene, TUB3; the combination of tub1-1 and a tub3 null mutation is inviable in haploids. Through further application of this method, new mutations in TUB2 and TUB3 were isolated as unlinked noncomplementers of tub1-1. The noncomplementation between tub1 and tub2 mutations is gene specific and allele specific, suggesting that the phenotype is due to an interaction at the protein level. We conclude that isolation of unlinked noncomplementing mutations is likely to be a generally useful method for isolating mutations in interacting gene products.

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