title: Mutations that affect the translation efficiency of Tn9-derived cat gene in Bacillus subtilis.
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.
authors.forename: C K
authors.initials: CK
authors.lastname: Lin
authors.forename: D S
authors.initials: DS
authors.lastname: Goldfarb
authors.forename: R H
authors.initials: RH
authors.lastname: Doi
authors.forename: R L
authors.initials: RL
authors.lastname: Rodriguez
mesh_terms: D000123:Acetyltransferases / Q000235:genetics*; D001412:Bacillus subtilis / Q000235:genetics*; D001665:Binding Sites; D015500:Chloramphenicol O-Acetyltransferase; D002872:Chromosome Deletion; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000235:genetics; D005786:Gene Expression Regulation; D005798:Genes, Bacterial; D009154:Mutation; D009690:Nucleic Acid Conformation; D010442:Peptide Chain Initiation, Translational; D011401:Promoter Regions, Genetic; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000378:metabolism; D012270:Ribosomes / Q000378:metabolism*
chemical_list: D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D012333:RNA, Messenger; D000123:Acetyltransferases; D015500:Chloramphenicol O-Acetyltransferase