Mutations that affect the translation efficiency of Tn9-derived cat gene in Bacillus subtilis

C. K. Lin, D. S. Goldfarb, R. H. Doi, R. L. Rodriguez

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

We have isolated two spontaneous mutations that increase the expression of the Tn9-derived cat gene in Bacillus subtilis. These mutations, which appear to affect initiation of translation of chloramphenicol acetyltransferase (CAT; acetyl-CoA:chloramphenicol 3-O-acetyltransferase, EC 2.3.1.28), consist of a tandem duplication and triplication of a 55-base-pair sequence located at the 5' end of cat. Included in the repeated sequence are the Shine-Dalgarno site, initiation codon, and a region of dyad symmetry located within the structural portion of the cat gene. A striking feature of the mutated initiation sites is their potential to form stem-loop structures at the 5' end of the cat messenger RNA. Within the single-stranded loops of these structures are the ribosome binding site and initiation codon for the cat gene. It appears that the Gram-negative cat translation initiation site has mutated to permit efficient utilization in B. subtilis without directly affecting Shine-Dalgarno sequence homology. This report suggests that secondary structure in the vicinity of the Shine-Dalgarno site can exert a strong positive influence on the initiation of translation in B. subtilis.

Original languageEnglish (US)
Pages (from-to)173-177
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume82
Issue number1
StatePublished - 1985

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Bacillus subtilis
Cats
Mutation
Chloramphenicol O-Acetyltransferase
Genes
Initiator Codon
Sequence Homology
Ribosomes
Base Pairing
Binding Sites
Messenger RNA

ASJC Scopus subject areas

  • General
  • Genetics

Cite this

Mutations that affect the translation efficiency of Tn9-derived cat gene in Bacillus subtilis. / Lin, C. K.; Goldfarb, D. S.; Doi, R. H.; Rodriguez, R. L.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 82, No. 1, 1985, p. 173-177.

Research output: Contribution to journalArticle

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AB - We have isolated two spontaneous mutations that increase the expression of the Tn9-derived cat gene in Bacillus subtilis. These mutations, which appear to affect initiation of translation of chloramphenicol acetyltransferase (CAT; acetyl-CoA:chloramphenicol 3-O-acetyltransferase, EC 2.3.1.28), consist of a tandem duplication and triplication of a 55-base-pair sequence located at the 5' end of cat. Included in the repeated sequence are the Shine-Dalgarno site, initiation codon, and a region of dyad symmetry located within the structural portion of the cat gene. A striking feature of the mutated initiation sites is their potential to form stem-loop structures at the 5' end of the cat messenger RNA. Within the single-stranded loops of these structures are the ribosome binding site and initiation codon for the cat gene. It appears that the Gram-negative cat translation initiation site has mutated to permit efficient utilization in B. subtilis without directly affecting Shine-Dalgarno sequence homology. This report suggests that secondary structure in the vicinity of the Shine-Dalgarno site can exert a strong positive influence on the initiation of translation in B. subtilis.

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