PMID:6328492

Citation	Chu, FK, Maley, GF, Maley, F and Belfort, M (1984) Intervening sequence in the thymidylate synthase gene of bacteriophage T4. Proc. Natl. Acad. Sci. U.S.A. 81:3049-53
Abstract	The continuous sequence of 2.3 kilobases in a 3-kilobase DNA fragment encoding the structural gene for coliphage T4 thymidylate synthase (5,10-methylenetetrahydrofolate:dUMP C-methyltransferase, EC 2.1.1.45) was determined by using the M13 dideoxy chain-termination method. From the coding information within this gene and that provided by sequence analysis of selected CNBr peptides from the protein product, the primary structure of T4 thymidylate synthase was determined. The most significant finding of these studies is the presence of a 1017-base-pair interruption two-thirds of the way through the nucleotide sequence of the structural gene. The 5'- and 3'-terminal ends of this intron are demarcated by an apparent stop and start codon, respectively. The corresponding methionine preceding the second coding region of the synthase is not incorporated into the final protein product. Structural evidence confirming the presence of the intervening sequence in the phage genome was obtained by restriction and hybridization analysis. Support for the presence of the intron was also obtained at the functional level by enzyme expression studies using selected td gene fragments. This work also confirms the findings of Purohit and Mathews [ Purohit , S. & Mathews , C. K. (1983) Fed. Proc. Fed. Am. Soc. Exp. Biol. 42, 1759], which reveal that the termination codon for the dihydrofolate reductase gene and the triplet initiating thymidylate synthase overlap by a four-base stretch, A-T-G-A. The implications of this unusual gene arrangement are discussed.
Links	PubMed PMC345218
Keywords	Amino Acid Sequence; Base Sequence; Cloning, Molecular; DNA Restriction Enzymes; Escherichia coli/enzymology; Escherichia coli/genetics; Genes; Genes, Viral; Methyltransferases/genetics; Plasmids; T-Phages/enzymology; T-Phages/genetics; Thymidylate Synthase/genetics
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
BPT4:TYSY	Contributes to	GO:0004799: thymidylate synthase activity	ECO:0000315:		F		Figure 1: Various restriction enzymes used to make cuts in the thymidylate synthase gene, and clones were generated and analyzed from the fragments.	complete CACAO 12079

References

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