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Rat C1-tetrahydrofolate synthase. cDNA isolation, tissue-specific levels of the mRNA, and expression of the protein in yeast.
J Biol Chem. 1990 May 15; 265(14):7907-13.JB

Abstract

We have isolated and characterized cDNA clones encoding rat cytoplasmic C1-tetrahydrofolate (H4folate) synthase. In eukaryotes, this enzyme is trifunctional and contains the activities of 10-formyl-H4folate synthetase, 5,10-methenyl-H4folate cyclohydrolase, and 5,10-methylene-H4folate dehydrogenase. The deduced sequence of the 935-amino acid open reading frame contained exact matches to NH2-terminal (15 residues) and internal (residues 436-450) peptide sequences obtained from the purified enzyme. The amino acid sequence derived from the rat cDNA shows extensive homology to analogous proteins from bacterial, yeast, and mammalian sources. We have used the cDNA to determine the steady-state levels of the mRNA in various rat tissues and have found that gene expression is regulated in a tissue-specific manner. Transcript levels are highest in kidney and liver with liver transcripts reduced about 30% relative to those found in kidney. Brain, heart, testis, lung and skeletal muscle display even lower transcript levels; reductions range from 70 to 80% of transcript levels found in kidney. Comparison to the levels of enzyme in these tissues allows us to conclude that pretranslational events predominate in the tissue-specific expression. The rat enzyme has been functionally expressed in Saccharomyces cerevisiae as evidenced by its capacity to complement a chromosomal deletion of ADE3, the yeast gene encoding cytoplasmic C1-H4folate synthase.

Authors+Show Affiliations

Clayton Foundation Biochemical Institute, Department of Chemistry, University of Texas, Austin 78712.No affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

2186031

Citation

Thigpen, A E., et al. "Rat C1-tetrahydrofolate Synthase. cDNA Isolation, Tissue-specific Levels of the mRNA, and Expression of the Protein in Yeast." The Journal of Biological Chemistry, vol. 265, no. 14, 1990, pp. 7907-13.
Thigpen AE, West MG, Appling DR. Rat C1-tetrahydrofolate synthase. cDNA isolation, tissue-specific levels of the mRNA, and expression of the protein in yeast. J Biol Chem. 1990;265(14):7907-13.
Thigpen, A. E., West, M. G., & Appling, D. R. (1990). Rat C1-tetrahydrofolate synthase. cDNA isolation, tissue-specific levels of the mRNA, and expression of the protein in yeast. The Journal of Biological Chemistry, 265(14), 7907-13.
Thigpen AE, West MG, Appling DR. Rat C1-tetrahydrofolate Synthase. cDNA Isolation, Tissue-specific Levels of the mRNA, and Expression of the Protein in Yeast. J Biol Chem. 1990 May 15;265(14):7907-13. PubMed PMID: 2186031.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Rat C1-tetrahydrofolate synthase. cDNA isolation, tissue-specific levels of the mRNA, and expression of the protein in yeast. AU - Thigpen,A E, AU - West,M G, AU - Appling,D R, PY - 1990/5/15/pubmed PY - 1990/5/15/medline PY - 1990/5/15/entrez SP - 7907 EP - 13 JF - The Journal of biological chemistry JO - J. Biol. Chem. VL - 265 IS - 14 N2 - We have isolated and characterized cDNA clones encoding rat cytoplasmic C1-tetrahydrofolate (H4folate) synthase. In eukaryotes, this enzyme is trifunctional and contains the activities of 10-formyl-H4folate synthetase, 5,10-methenyl-H4folate cyclohydrolase, and 5,10-methylene-H4folate dehydrogenase. The deduced sequence of the 935-amino acid open reading frame contained exact matches to NH2-terminal (15 residues) and internal (residues 436-450) peptide sequences obtained from the purified enzyme. The amino acid sequence derived from the rat cDNA shows extensive homology to analogous proteins from bacterial, yeast, and mammalian sources. We have used the cDNA to determine the steady-state levels of the mRNA in various rat tissues and have found that gene expression is regulated in a tissue-specific manner. Transcript levels are highest in kidney and liver with liver transcripts reduced about 30% relative to those found in kidney. Brain, heart, testis, lung and skeletal muscle display even lower transcript levels; reductions range from 70 to 80% of transcript levels found in kidney. Comparison to the levels of enzyme in these tissues allows us to conclude that pretranslational events predominate in the tissue-specific expression. The rat enzyme has been functionally expressed in Saccharomyces cerevisiae as evidenced by its capacity to complement a chromosomal deletion of ADE3, the yeast gene encoding cytoplasmic C1-H4folate synthase. SN - 0021-9258 UR - https://www.unboundmedicine.com/medline/citation/2186031/Rat_C1_tetrahydrofolate_synthase__cDNA_isolation_tissue_specific_levels_of_the_mRNA_and_expression_of_the_protein_in_yeast_ L2 - http://www.jbc.org/cgi/pmidlookup?view=long&pmid=2186031 DB - PRIME DP - Unbound Medicine ER -