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Single-stranded DNA binding activity of C1-tetrahydrofolate synthase enzymes.
J Biol Chem. 1993 Nov 15; 268(32):23792-8.JB

Abstract

In eukaryotes C1-5,6,7,8-tetrahydrofolate (THF) synthase is a trifunctional enzyme that catalyzes the interconversion of reduced forms of folate to supply activated one-carbon units required for a variety of metabolic pathways. The enzymatic activities include 10-formyl-THF synthetase (EC 6.3.4.3), 5,10-methenyl-THF cyclohydrolase (EC 3.5.4.9), and 5,10-methylene-THF dehydrogenase (EC 1.5.1.5). In bacteria separate, monofunctional or bifunctional polypeptides catalyze the same reactions. We have purified C1-THF synthase from the fission yeast Schizosaccharomyces pombe and found its physical and enzymatic properties similar to those of other eukaryotic C1-THF synthase enzymes. Unexpectedly, the S. pombe enzyme bound strongly (Keq = 100 pM) to single-stranded DNA, but not to double-stranded DNA or to RNA. The binding was sequence-independent, apparently not cooperative, and not detectably inhibited by C1-THF synthase substrates or cofactors. Trifunctional cytoplasmic enzyme from Saccharomyces cerevisiae and monofunctional (synthetase) enzyme from Clostridium acidiurici also bound tightly to single-stranded DNA, while bifunctional (dehydrogenase and cyclohydrolase) enzyme from Escherichia coli did not, suggesting that single-stranded DNA binding is a conserved function of the synthetase domain of C1-THF synthase enzymes.

Authors+Show Affiliations

Fred Hutchinson Cancer Research Center, Seattle, Washington 98104.No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

8226914

Citation

Wahls, W P., et al. "Single-stranded DNA Binding Activity of C1-tetrahydrofolate Synthase Enzymes." The Journal of Biological Chemistry, vol. 268, no. 32, 1993, pp. 23792-8.
Wahls WP, Song JM, Smith GR. Single-stranded DNA binding activity of C1-tetrahydrofolate synthase enzymes. J Biol Chem. 1993;268(32):23792-8.
Wahls, W. P., Song, J. M., & Smith, G. R. (1993). Single-stranded DNA binding activity of C1-tetrahydrofolate synthase enzymes. The Journal of Biological Chemistry, 268(32), 23792-8.
Wahls WP, Song JM, Smith GR. Single-stranded DNA Binding Activity of C1-tetrahydrofolate Synthase Enzymes. J Biol Chem. 1993 Nov 15;268(32):23792-8. PubMed PMID: 8226914.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Single-stranded DNA binding activity of C1-tetrahydrofolate synthase enzymes. AU - Wahls,W P, AU - Song,J M, AU - Smith,G R, PY - 1993/11/15/pubmed PY - 1993/11/15/medline PY - 1993/11/15/entrez SP - 23792 EP - 8 JF - The Journal of biological chemistry JO - J. Biol. Chem. VL - 268 IS - 32 N2 - In eukaryotes C1-5,6,7,8-tetrahydrofolate (THF) synthase is a trifunctional enzyme that catalyzes the interconversion of reduced forms of folate to supply activated one-carbon units required for a variety of metabolic pathways. The enzymatic activities include 10-formyl-THF synthetase (EC 6.3.4.3), 5,10-methenyl-THF cyclohydrolase (EC 3.5.4.9), and 5,10-methylene-THF dehydrogenase (EC 1.5.1.5). In bacteria separate, monofunctional or bifunctional polypeptides catalyze the same reactions. We have purified C1-THF synthase from the fission yeast Schizosaccharomyces pombe and found its physical and enzymatic properties similar to those of other eukaryotic C1-THF synthase enzymes. Unexpectedly, the S. pombe enzyme bound strongly (Keq = 100 pM) to single-stranded DNA, but not to double-stranded DNA or to RNA. The binding was sequence-independent, apparently not cooperative, and not detectably inhibited by C1-THF synthase substrates or cofactors. Trifunctional cytoplasmic enzyme from Saccharomyces cerevisiae and monofunctional (synthetase) enzyme from Clostridium acidiurici also bound tightly to single-stranded DNA, while bifunctional (dehydrogenase and cyclohydrolase) enzyme from Escherichia coli did not, suggesting that single-stranded DNA binding is a conserved function of the synthetase domain of C1-THF synthase enzymes. SN - 0021-9258 UR - https://www.unboundmedicine.com/medline/citation/8226914/Single_stranded_DNA_binding_activity_of_C1_tetrahydrofolate_synthase_enzymes_ L2 - http://www.jbc.org/cgi/pmidlookup?view=long&pmid=8226914 DB - PRIME DP - Unbound Medicine ER -