Role in translation of a triple tandemly repeated sequence in the 5′-untranslated region of human thymidylate synthase mRNA

Sumiko Kaneda, Keiichi Takeishi, Dai Ayusawa, Kimiko Shimizu, Takeshi Seno, Sidney Altman

Research output: Contribution to journalArticle

135 Citations (Scopus)

Abstract

A triple tandem repeat (TTR) consisting of 90 nucleotides exists immediately upstream of the ATG initiator codon in human thymidylate synthase (TS) cDNA (pcHTS-1). To investigate the role of the TTR in the expression of the TS cDNA, we used pcHTS-1 to construct mutant cDNA clones in which part of the TTR was deleted or an additional element was inserted. The mutant cDNA plasmid was introduced into murine TS-negat1ve mutant cells and the relative translation efficiencies of the mutant cDNAs were determined by measuring the transient expression of TS activity and the amount of TS mRNA transcribed. The translation efficiency in transient expression of the mutants was increased by deletions covering all the first two repeated elements, and the part of the third closest to the ATG initiator codon, but was not affected by deletions of only parts of the first two repeated elements at the 5′ end. The translation efficiency was also not affected by insertion of an additional repeated element Into the TTR. These results suggest that the first two repeated elements at the 5′ end both have inhibitory effects on translation of the TS mRNA, probably due to the unique structural feature of this element.

Original languageEnglish (US)
Pages (from-to)1259-1270
Number of pages12
JournalNucleic Acids Research
Volume15
Issue number3
DOIs
StatePublished - Feb 11 1987
Externally publishedYes

Fingerprint

Thymidylate Synthase
5' Untranslated Regions
CDNA
Mutant
Messenger RNA
Tandem Repeat Sequences
Complementary DNA
Initiator Codon
Deletion
Nucleotides
Clone
Insertion
Immediately
Covering
Human
cDNA
Plasmids
Clone Cells
Cell

ASJC Scopus subject areas

  • Statistics, Probability and Uncertainty
  • Applied Mathematics
  • Health, Toxicology and Mutagenesis
  • Toxicology
  • Genetics(clinical)
  • Genetics

Cite this

Role in translation of a triple tandemly repeated sequence in the 5′-untranslated region of human thymidylate synthase mRNA. / Kaneda, Sumiko; Takeishi, Keiichi; Ayusawa, Dai; Shimizu, Kimiko; Seno, Takeshi; Altman, Sidney.

In: Nucleic Acids Research, Vol. 15, No. 3, 11.02.1987, p. 1259-1270.

Research output: Contribution to journalArticle

Kaneda, Sumiko ; Takeishi, Keiichi ; Ayusawa, Dai ; Shimizu, Kimiko ; Seno, Takeshi ; Altman, Sidney. / Role in translation of a triple tandemly repeated sequence in the 5′-untranslated region of human thymidylate synthase mRNA. In: Nucleic Acids Research. 1987 ; Vol. 15, No. 3. pp. 1259-1270.
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