Mutations in the gene encoding the 34 kDa subunit of yeast replication protein A cause defective S phase progression
1995
Santocanale, C. | Neecke, H. | Longhese, M.P. | Lucchini, G. | Plevani, P.
The in vivo function of the 34 kDa subunit of yeast replication protein A (RPA), encoded by the RFA2 gene, has been studied by analyzing the effect of Rpa34 depletion and by producing and characterizing rfa2 temperature sensitive mutants, We show that unbalanced stoichiometry of the RPA subunits does not affect cell growth and cell cycle progression until the level of Rpa34 becomes rate-limiting, at which point cells arrest with a late S/G2 DNA content. Rpa34 is involved in DNA replication in vivo, since rfa2 ts mutants are defective in S phase progression and ARS plasmid stability, and rfa2 pol1 double mutants are non-viable. Moreover, when shifted to the restrictive temperature, about 50% of the rfa2 mutant cells rapidly die while traversing the S phase and the surviving cells arrest in late S/G2 at the RAD9 checkpoint. Finally, rfa2 mutant cells have a mutator and hyper-recombination phenotype and are more sensitive to hydroxyurea and methyl-methane-sulfonate than wild-type cells.
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