Skip to main content
PMC home page
Genetics logoLink to Genetics
. 1993 Feb;133(2):183–191. doi: 10.1093/genetics/133.2.183

Conditional Mutations in the Yeast DNA Primase Genes Affect Different Aspects of DNA Metabolism and Interactions in the DNA Polymerase α-Primase Complex

M P Longhese 1, L Jovine 1, P Plevani 1, G Lucchini 1
PMCID: PMC1205309  PMID: 8436268

Abstract

Different pri1 and pri2 conditional mutants of Saccharomyces cerevisiae altered, respectively, in the small (p48) and large (p58) subunits of DNA primase, show an enhanced rate of both mitotic intrachromosomal recombination and spontaneous mutation, to an extent which is correlated with the severity of their defects in cell growth and DNA synthesis. These effects might be attributable to the formation of nicked and gapped DNA molecules that are substrates for recombination and error-prone repair, due to defective DNA replication in the primase mutants. Furthermore, pri1 and pri2 mutations inhibit sporulation and affect spore viability, with the unsporulated mutant cells arresting with a single nucleus, suggesting that DNA primase plays a critical role during meiosis. The observation that all possible pairwise combinations of two pri1 and two pri2 alleles are lethal provides further evidence for direct interaction of the primase subunits in vivo. Immunopurification and immunoprecipitation studies on wild-type and mutant strains suggest that the small subunit has a major role in determining primase activity, whereas the large subunit directly interacts with DNA polymerase α, and either mediates or stabilizes association of the p48 polypeptide in the DNA polymerase α-primase complex.

Full Text

The Full Text of this article is available as a PDF (3.3 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aguilera A., Klein H. L. Genetic control of intrachromosomal recombination in Saccharomyces cerevisiae. I. Isolation and genetic characterization of hyper-recombination mutations. Genetics. 1988 Aug;119(4):779–790. doi: 10.1093/genetics/119.4.779. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brooks M., Dumas L. B. DNA primase isolated from the yeast DNA primase-DNA polymerase complex. Immunoaffinity purification and analysis of RNA primer synthesis. J Biol Chem. 1989 Feb 25;264(6):3602–3610. [PubMed] [Google Scholar]
  3. Budd M. E., Wittrup K. D., Bailey J. E., Campbell J. L. DNA polymerase I is required for premeiotic DNA replication and sporulation but not for X-ray repair in Saccharomyces cerevisiae. Mol Cell Biol. 1989 Feb;9(2):365–376. doi: 10.1128/mcb.9.2.365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Foiani M., Santocanale C., Plevani P., Lucchini G. A single essential gene, PRI2, encodes the large subunit of DNA primase in Saccharomyces cerevisiae. Mol Cell Biol. 1989 Jul;9(7):3081–3087. doi: 10.1128/mcb.9.7.3081. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Game J. C., Johnston L. H., von Borstel R. C. Enhanced mitotic recombination in a ligase-defective mutant of the yeast Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4589–4592. doi: 10.1073/pnas.76.9.4589. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hartwell L. H., Smith D. Altered fidelity of mitotic chromosome transmission in cell cycle mutants of S. cerevisiae. Genetics. 1985 Jul;110(3):381–395. doi: 10.1093/genetics/110.3.381. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Lehman I. R., Kaguni L. S. DNA polymerase alpha. J Biol Chem. 1989 Mar 15;264(8):4265–4268. [PubMed] [Google Scholar]
  8. Lucchini G., Biraghi A., Carbone M. L., de Scrilli A., Magni G. E. Effect of mutation in the aromatic amino acid pathway on sporulation of Saccharomyces cerevisiae. J Bacteriol. 1978 Oct;136(1):55–62. doi: 10.1128/jb.136.1.55-62.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Lucchini G., Brandazza A., Badaracco G., Bianchi M., Plevani P. Identification of the yeast DNA polymerase I gene with antibody probes. Curr Genet. 1985;10(4):245–252. doi: 10.1007/BF00365620. [DOI] [PubMed] [Google Scholar]
  10. Lucchini G., Muzi Falconi M., Pizzagalli A., Aguilera A., Klein H. L., Plevani P. Nucleotide sequence and characterization of temperature-sensitive pol1 mutants of Saccharomyces cerevisiae. Gene. 1990 May 31;90(1):99–104. doi: 10.1016/0378-1119(90)90444-v. [DOI] [PubMed] [Google Scholar]
  11. Plevani P., Foiani M., Valsasnini P., Badaracco G., Cheriathundam E., Chang L. M. Polypeptide structure of DNA primase from a yeast DNA polymerase-primase complex. J Biol Chem. 1985 Jun 10;260(11):7102–7107. [PubMed] [Google Scholar]
  12. Plevani P., Francesconi S., Lucchini G. The nucleotide sequence of the PRI1 gene related to DNA primase in Saccharomyces cerevisiae. Nucleic Acids Res. 1987 Oct 12;15(19):7975–7989. doi: 10.1093/nar/15.19.7975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Prussak C. E., Almazan M. T., Tseng B. Y. Mouse primase p49 subunit molecular cloning indicates conserved and divergent regions. J Biol Chem. 1989 Mar 25;264(9):4957–4963. [PubMed] [Google Scholar]
  14. Trueheart J., Boeke J. D., Fink G. R. Two genes required for cell fusion during yeast conjugation: evidence for a pheromone-induced surface protein. Mol Cell Biol. 1987 Jul;7(7):2316–2328. doi: 10.1128/mcb.7.7.2316. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Tsurimoto T., Melendy T., Stillman B. Sequential initiation of lagging and leading strand synthesis by two different polymerase complexes at the SV40 DNA replication origin. Nature. 1990 Aug 9;346(6284):534–539. doi: 10.1038/346534a0. [DOI] [PubMed] [Google Scholar]

Articles from Genetics are provided here courtesy of Oxford University Press

RESOURCES