Prior preparation of N(5),N(10)-methylenetetrahydrofolate from dl-tetrahydrofolate and [(14)C]formaldehyde resulted in an improved assay for thymidylate synthase. Although preparations from tobacco seedlings and cotton root tips (0.25 centimeter) were inconsistent with respect to enzyme activity, extracts from actively growing cell cultures of Chlamydomonas, Nicotiana hybrid callus, Pinus callu...

The effects of the lipid-soluble dihydrofolate reductase inhibitor, trimetrexate, on the inhibition of thymidylate biosynthesis as a result of perturbation in cellular folate pools in H35 hepatoma cells in vitro has been investigated. Exposure of the cultures to increasing concentrations of trimetrexate between 2 and 20 nM causes a marked reduction in de novo thymidylate biosynthesis and a conc...

The DNA nucleotide thymidylate is synthesized by the enzyme thymidylate synthase, which catalyzes the reductive methylation of deoxyuridylate using the cofactor methylene-tetrahydrofolate (CH(2)H(4)folate). Most organisms, including humans, rely on the thyA- or TYMS-encoded classic thymidylate synthase, whereas, certain microorganisms, including all Rickettsia and other pathogens, use an altern...

The rTSbeta protein has been hypothesized to synthesize signaling molecules that can down-regulate thymidylate synthase. These molecules share biological and chemical properties with acyl-homoserine lactones (AHL), suggesting some AHLs might act as rTS signaling mimics and down-regulate thymidylate synthase. We have determined that the AHL, 3-oxododecanoyl homoserine lactone (3-oxo-C12-(L)-HSL)...

The effects of 5,8-dideazaisopteroylglutamate (A/-[p-{[(2amino-4-hydroxy-6-quinazolinyl)amino]methyl}benzoyl]-Lglutamic acid) (IAHQ) on thymidylate synthase and dihydrofolate reducÃ-ase have been examined as a biochemical basis for the anticancer activity of IAHQ in several tumor systems. IAHQ, when administered at 85 mg/kg on Days 2 and 10 after tumor implantation, delayed the growth of methot...

The de novo thymidylate biosynthetic pathway in mammalian cells translocates to the nucleus for DNA replication and repair and consists of the enzymes serine hydroxymethyltransferase 1 and 2α (SHMT1 and SHMT2α), thymidylate synthase, and dihydrofolate reductase. In this study, we demonstrate that this pathway forms a multienzyme complex that is associated with the nuclear lamina. SHMT1 or SHMT2...

Folate analogs that inhibit dihydrofolate reductase result in only partial interconversion of tetrahydrofolate cofactors to dihydrofolate with preservation of the major portion of reduced cellular folate cofactors in L1210 leukemia cells. One possible explanation for this phenomenon is that low levels of dihydrofolate polyglutamates that accumulate in the presence of antifolates block thymidyla...