Summary
Seven temperature-sensitive mutants have been isolated in Saccharomyces cerevisiae which show a reproducible defect in DNA synthesis at the restrictive temperature. One of these is allelic with rnal1 (Hartwell et al., 1970) but the remaining mutants define six complementation groups and probably represent six different genes. The gene symbol dds (for depressed DNA synthesis) is proposed.
At the restrictive temperature, rnal1-2, dds2-1 and dds6-1 show a rapid and almost total cessation of DNA and RNA synthesis, whilst protein synthesis continues for several hours. The remaining dds mutants show a reduced rate of DNA synthesis from the time of temperature shift (dds1, dds3, dds4) or a cessation of DNA synthesis at a later time (dds5). In some cases, RNA synthesis is affected concomitantly with, or soon after, the depression in DNA synthesis. Possible reasons for the phenotypes of these mutants, and for the relative absence of yeast mutants which are unambiguously and specifically affected in DNA synthesis, are discussed.
In addition, we report the isolation of seven new alleles of known cdc genes and ten new mutants with a cell cycle phenotype that complement those already known.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bisson, L., Thorner, J.: Thymidine 5′-monophosphate-requiring mutants of Saccharomyces cerevisiae are deficient in thymidylate synthatase. J. Bact. 132, 44–50 (1977)
Byers, B., Goetsch, L.: Duplication of spindle plaques and integration of the yeast cell cycle. Cold Spr. Harb. Symp. quant. Biol. 38, 123–131 (1973)
Campbell, J.L., Soll, L., Richardson, C.C.: Isolation and partial characterisation of a mutant of E. coli deficient in polII. Proc. nat. Acad. Sci. (Wash.) 69, 2090–2094. (1972)
Flickinger, R.A.: Replication-dependent transcription in eukaryotes. Int. J. Biochem. 7, 85–93 (1976)
Game, J.: Yeast cell cycle mutant cdc21 in a temperature-sensitive thymidylate auxotroph. Molec. gen. Genet. 146, 313–315 (1976)
Gefter, M.: DNA replication. Ann. Rev. Biochem. 44, 45–78 (1975)
Gefter, M., Hirota, Y., Kornberg, T., Wechsler, J., Barnoux, L.: Analysis of DNA polymerase II and III in mutants of E. coli thermosensitive for DNA synthesis. Proc. nat. Acad. Sci. (Wash.) 68, 3150–3154 (1971)
Hartwell, L.H.: Macromolecule synthesis in temperature-sensitive mutants of yeast. J. Bact. 93, 1662–1670 (1967)
Hartwell, L.H.: Genetic control of the cell cycle in yeast. II. Genes controlling DNA replication and its initiation. J. molec. Biol. 59, 183–194 (1971)
Hartwell, L.H.: Three additional genes required for DNA synthesis in S. cerevisiae. J. Bact. 115, 966–974 (1973)
Hartwell, L.H.: Saccharomyces cerevisiae cell cycle. Bact. Rev. 38, 164–198 (1974)
Hartwell, L.H., McLaughlin, C.S., Warner, J.R.: Identification of ten genes that control ribosome formation in yeast. Molec. gen. Genet. 109, 42–56 (1970)
Hartwell, L.H., Mortimer, R.K., Culotti, J., Culotti, M.: Genetic control of the cell division cycle in yeast. V. Genetic analysis of cdc mutants. Genetics 74, 267–286 (1973)
Hereford, L.M., Hartwell, L.H.: Sequential gene function in the initiation of S. cerevisiae DNA synthesis. J. molec. Biol. 84, 445–461 (1974)
Hutchison, H., Hartwell, L.H., McLaughlin, C.S.: A temperaturesensitive yeast mutant defective in RNA production. J. Bact. 99, 807–814 (1969)
Johnston, L.H.: The use of a novel plate assay in a search for yeast mutants defective in deoxyribonucleases. Molec. gen. Genet. 152, 219–222 (1977)
Johnston, L.H., Bonhoeffer, F., Symmons, P.: Molecular principles and the enzymatic machinery of DNA replication. In: Cell biology: A comprehensive treatise, Vol. 2 New York: Academic Press 1977 (in press)
Kornberg, A.: DNA synthesis. San Francisco: W.H. Freeman and Co., 1974
Lucia, P. de, Cairns, J.: Isolation of an E. coli strain with a mutation affecting DNA polymerase. Nature (Lond.) 224, 1164–1166 (1969)
McClary, D., Nulty, W.L., Miller, G.R.: Effect of potassium versus sodium in the sporulation of Saccharomyces. J. Bact. 78, 362–380 (1959)
Mortimer, R.K., Hawthorne, D.C.: Yeast genetics. In: The yeasts (Rose, A.H., Harrison, J.S., eds.). London: Academic Press 1969
Mortimer, R.K., Hawthorne, D.C.: Genetic mapping in Saccharomyces. IV. Mapping of temperature-sensitive genes and use of disomic strains in localising genes. Genetics 74, 33–54 (1973)
Nusslein, V., Otto, B., Bonhoeffer, F., Schaller, H.: Function of DNA polymerase III in DNA replication. Nature (Lond.) 234, 285–286 (1971)
Plischke, M.E., von Borstel, R.C., Mortimer, R.K., Cohn, W.E.: Genetic markers and associated gene products in S. cerevisiae. In: Handbook of biochemistry and molecular biology, 3rd ed. Nucleic acids, Vol. II (Fasman, G.D., ed.), pp. 767–832 Cleveland, Ohio: Chemical Rubber Co. Press (1976)
Thonart, P., Bechet, J., Hilgar, F., Burny, A.: Thermosensitive mutations affecting ribonucleic acid polymerases in S. cerevisiae. J. Bact. 125, 25–32 (1976)
Unrau, P., Hollyday, R.: A search of temperature-sensitive mutants of U. maydis blocked in DNA synthesis. Genet. Res. Camb. 15, 157–169 (1970)
Author information
Authors and Affiliations
Additional information
Communicated by W. Gajewski
Rights and permissions
About this article
Cite this article
Johnston, L.H., Game, J.C. Mutants of yeast with depressed DNA synthesis. Molec. Gen. Genet. 161, 205–214 (1978). https://doi.org/10.1007/BF00274189
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00274189