Thymidylate Synthase Heterogeneity Assessed by Monoclonal Antibodies

  title={Thymidylate Synthase Heterogeneity Assessed by Monoclonal Antibodies},
  author={B Gołos and Elżbieta Wałajtys-Rode and Anna Porębska and Joanna Cieśla and M Dabrowska and Zbigniew Zieliński and Wojciech Rode},
Thymidylate synthase (TS; EC catalyzes N5,10-methylenetetrahydrofolate-dependent methylation of dUMP. Purified preparations of the enzyme from regenerating rat liver, L1210 cell and Trichinella spiralis muscle larvae, and recombinant rat hepatoma and mouse L1210 thymidylate synthases, analyzed with the use of SDS Polyacrylamide gel electrophoresis, show heterogeneity, reflected by an additional distinct band (Figure 1; cf. ref. 1), located too close to the main band to enable… Expand
The effect of Arg209 to Lys mutation in mouse thymidylate synthase.
It is documented that mouse thymidylate synthase R209, similar to the corresponding L. casei R218, is essential for both dUMP binding and enzyme reaction. Expand
Altered mouse leukemia L1210 thymidylate synthase, associated with cell resistance to 5-fluoro-dUrd, is not mutated but rather reflects posttranslational modification.
Direct sequencing of products obtained from RT-PCR proved both nucleotide sequences to be identical to the mouse thymidylate synthase coding sequence published earlier, suggesting that the altered properties of TSr are caused by a factor different than protein mutation, presumably posttranslational modification. Expand
Binding and repression of translation of the cognate mRNA by Trichinella spiralis thymidylate synthase differ from the corresponding interactions of the human enzyme.
It is shown that mRNA binding requires a methionine or an adduct at the N-terminus of TS, but that the translational repression effect does not, and that chicken liver dihydrofolate reductase, which is incapable of binding to T. spiralis TS mRNA, repressed the translation of TS. Expand
Developmental arrest in Caenorhabditis elegans dauer larvae causes high expression of enzymes involved in thymidylate biosynthesis, similar to that found in Trichinella muscle larvae
High thymidylate synthase expression in developmentally arrested larvae, demonstrated also at the mRNA and protein levels, is in agreement with a global cell cycle arrest of dauer larvae and indicates this unusual cell cycle regulation pattern can be shared by developmentally arrests larvae of C. elegans and the two Trichnella species. Expand


Thymidylate synthases from Hymenolepis diminuta and regenerating rat liver: purification, properties, and inhibition by substrate and cofactor analogues.
Comparative studies of thymidylate synthases, isolated from the tapeworm, Hymenolepis diminuta, and regenerating liver of its host, rat, aimed at a possibility of specific inhibition of theExpand
Intracellular Location of Thymidylate Synthase and Its State of Phosphorylation*
Findings suggest that this enzyme’s contribution to the cell cycle may be more complex than believed previously, and that the enzyme appears to be a phosphoprotein, similar to that found for other proteins associated with cell division and signal transduction. Expand
Trichinella spiralis thymidylate synthase: developmental pattern, isolation, molecular properties, and inhibition by substrate and cofactor analogues.
Thymidylate synthase specific activity was found to remain at a constant level in crude extracts from muscle larvae, isolated (1-15 months after infection) by pepsin-HCI digestion, as well as fromExpand
Isolation and expression of rat thymidylate synthase cDNA: phylogenetic comparison with human and mouse thymidylate synthases.
Two cDNA clones representing rat hepatoma thymidylate synthase (rTS) were isolated from a lambda ZAP II cDNA library using as a probe a fragment of the human TS cDNA, and it is suggested that mouse TS is closer to human TS phylogenetically than rTS. Expand
Mechanism of inhibition of mammalian tumor and other thymidylate synthases by N4-hydroxy-dCMP, N4-hydroxy-5-fluoro-dCMP, and related analogues.
Kinetic studies with purified enzyme from five sources demonstrated that addition of a 5-fluoro substituent to N4-OH-dCMP increased its affinity from 2- to 20-fold for the enzyme from different sources. Expand