Topic
dTMP kinase
About: dTMP kinase is a research topic. Over the lifetime, 43 publications have been published within this topic receiving 995 citations.
Papers
TL;DR: 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, and it is concluded that CDC8 is the structural gene for dTMP kinase, which catalyzes an essential step in DNA precursor biosynthesis.
Abstract: 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 has 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.
104 citations
TL;DR: A series of N(1)-(4-substituted-benzyl)-pyrimidines were synthesized as potential inhibitors of thymidine monophosphate kinase of Mycobacterium tuberculosis and key SAR parameters included the chain length substitution in para position of the benzyl ring, the functional group terminating the alkyl chain, and the substituent on the C-5 pyrimidine ring.
96 citations
TL;DR: The identification of the human dTMP kinase gene and expression of its product in the baculovirus expression system should facilitate study of the mechanism of gene regulation and its role in pyrimidine metabolism.
Abstract: dTMP kinase (E.C.2.7.4.9.) catalyzes the phosphorylation of dTMP to the corresponding diphosphate. This enzyme is essential for DNA synthesis in vivo and is an important intermediate enzyme in the pathway of many pyrimidine analog drugs. In this report, we describe the isolation of the human dTMP kinase gene by functional complementation of a Saccharomyces cerevisiae cell cycle mutant, cdc8. The cDNA sequence revealed an open reading frame that encodes a protein with the molecular weight of 23,806. The deduced protein sequence was compared to known dTMP kinase sequences from different organisms. Although functionally complementary and structurally conserved, expressed human dTMP kinase in yeast shows little enzymatic activity. In contrast, active human dTMP kinase can be expressed from the gene cloned into the baculovirus expression system, as evidenced by increased enzymatic activity by four- to five-fold. Unlike yeast dTMP kinase, human dTMP kinase does not contain a cysteine residue after the conserved glycine-rich loop, but its enzymatic activity is still affected by the sulfhydryl inhibitor, 5,5'-dithio-bis(2-nitrobenzoic acid) (DTNB). The levels of dTMP kinase mRNA and its enzymatic activity fluctuate during the cell cycle, peaking at the S phase. Thus, like Saccharomyces cerevisiae CDC8 (encoding dTMP kinase), the human homolog mRNA and enzymatic activity are also cell cycle regulated. We have also examined four neuroblastoma cell lines for dTMP kinase mRNA levels and its kinase activities, which appear to vary according to cell growth rate. Our results suggest that the expression of the dTMP kinase gene and its activity coincide with various stages of cell growth. The identification of the human dTMP kinase gene and expression of its product in the baculovirus expression system should facilitate study of the mechanism of gene regulation and its role in pyrimidine metabolism.
60 citations
TL;DR: In this article, the authors observed a periodism in the apparent level of dCMP deaminase during the growth of Chlorella pyrenoidosa and suggested that the enzymes on the dTTP biosynthetic pathway might limit the rate of DNA synthesis during the cell cycle.
42 citations
TL;DR: The results in this paper constitute direct evidence for the existence of multienzyme complexes of DNA synthesis in mammalian cells.
36 citations
Related Topics (5)
Performance Metrics
| Year | Papers |
|---|---|
| 2018 | 1 |
| 2013 | 1 |
| 2012 | 1 |
| 2010 | 1 |
| 2008 | 1 |
| 2007 | 1 |