Yeast gene CDC8 encodes thymidylate kinase and is complemented by herpes thymidine kinase gene TK

The herpes simplex virus type 1 thymidine kinase gene TK complements the defect in five temperature-sensitive mutants and in vitro constructed insertion and deletion mutants of the CDC8 gene of Saccharomyces cerevisiae. The herpes thymidine kinase enzyme acts as both a thymidine kinase and a thymidy...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1984-09, Vol.81 (18), p.5821-5825
Main Authors: Sclafani, Robert A., Fangman, Walton L.
Format: Article
Language:English
Subjects:
Alleles
Base Sequence
Biosynthesis
Diploidy
DNA
DNA replication
DNA Restriction Enzymes
Enzymes
Escherichia coli - enzymology
Escherichia coli - genetics
Genes
Genes, Fungal
Genes, Viral
Genetic Complementation Test
herpes simplex virus 1
Mutation
Nucleic acid precursors
Nucleoside-Phosphate Kinase - genetics
Phenotypes
Phosphotransferases - genetics
Plasmids
Saccharomyces cerevisiae
Saccharomyces cerevisiae - enzymology
Saccharomyces cerevisiae - genetics
Simplexvirus - enzymology
Simplexvirus - genetics
Temperature
Thymidine Kinase - genetics
Yeasts
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Summary:The herpes simplex virus type 1 thymidine kinase gene TK complements the defect in five temperature-sensitive mutants and in vitro constructed insertion and deletion mutants of the CDC8 gene of Saccharomyces cerevisiae. The herpes thymidine kinase enzyme acts as both a thymidine kinase and a thymidylate kinase (dTMP kinase). The latter activity is responsible for the cdc8 complementation since all thermosensitive cdc8 mutants are deficient in dTMP kinase activity at all temperatures. However, an intragenic revertant, cdc8-320, which was selected by demanding mitotic growth at the restrictive temperature, exhibits thermolabile dTMP kinase activity. We conclude that CDC8 is the structural gene for dTMP kinase, which catalyzes an essential step in DNA precursor biosynthesis. Previously, it had been shown that the DNA replication defect of cdc8 mutants could not be bypassed by the addition of deoxyribonucleoside triphosphates to permeabilized cells. This apparent discrepancy can be explained by hypothesizing a multiprotein yeast DNA replication complex containing the CDC8 protein.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.81.18.5821