Faithful chromosome transmission requires Spt4p, a putative regulator of chromatin structure in Saccharomyces cerevisiae

Munira A. Basrai, Jeffrey Kingsbury, Douglas Koshland, Forrest Spencer, Philip Hieter

Research output: Contribution to journalArticlepeer-review

Abstract

A chromosome transmission fidelity (ctf) mutant, s138, of Saccharomyces cerevisiae was identified by its centromere (CEN) transcriptional readthrough phenotype, suggesting perturbed kinetochore integrity in vivo. The gene complementing the s138 mutation was found to be identical to the S. cerevisiae SPT4 gene. The s138 mutation is a missense mutation in the second of four conserved cysteine residues positioned similarly to those of zinc finger proteins, and we henceforth refer to the mutation as spt4-138. Both spt4-138 and spt4Δ strains missegregate a chromosome fragment at the permissive temperature, are temperature sensitive for growth at 37°C, and upon a shift to the nonpermissive temperature show an accumulation of large budded cells, each with a nucleus. Previous studies suggest that Spt4p functions in a complex with Spt5p and Spt6p, and we determined that spt6-140 also causes missegregation of a chromosome fragment. Double mutants carrying spt4Δ2:: HIS3 and kinetochore mutation ndc10-42 or ctf13-30 show a synthetic conditional phenotype. Both spt4-138 and spt4Δ strains exhibit synergistic chromosome instability in combination with CEN DNA mutations and show in vitro defects in microtubule binding to minichromosomes. These results indicate that Spt4p plays a role in chromosome segregation. The results of in vivo genetic interactions with mutations in kinetochore proteins and CEN DNA and of in vitro biochemical assays suggest that Spt4p is important for kinetochure function.

Original languageEnglish (US)
Pages (from-to)2838-2847
Number of pages10
JournalMolecular and cellular biology
Volume16
Issue number6
DOIs
StatePublished - 1996

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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