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PMID: 1660923 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Programmed translational frameshifting and initiation at an AUU codon in gene expression of bacterial insertion sequence IS911.

Journal of molecular biology ·Vol. 222 ·No. 3 ·1991-12-05 ·Pages 465-77

Polard P, Prère MF, Chandler M, Fayet O

Abstract

The proteins expressed by insertion sequence IS911, a member of the widespread IS3 family of elements, have been analyzed. The results indicate that three major species are produced from two consecutive reading frames. A protein of Mr 11,500, ORFA, is synthesized from an upstream reading frame. A larger protein, ORFAB, uses the same initiation codon and is produced by a -1 programmed translational frameshift between orfA and a downstream frame, orfB, whose amino acid sequence shows significant homology with retroviral integrase proteins. The orfB frame is also expressed independently in two alternative forms: the first uses a rare AUU initiation codon in the orfB phase whereas the second appears to initiate in the orfA phase and is produced by a -1 frameshift mechanism similar to that used in ORFAB expression. A specific IS911 integration reaction using a minimal active junction composed of 51 base-pairs of the right inverted repeat and a flanking phase lambda sequence resembling a second end in inverted orientation has been developed to analyze the functions of these proteins by transcomplementation in vivo. The orfA and orfB frames are shown to be essential and production of ORFAB is shown to stimulate integration in this system, suggesting that this fusion protein is the IS911 transposase.

MeSH Terms
Amino Acid Sequence Base Sequence Cloning, Molecular Codon/genetics DNA Transposable Elements/genetics Enterobacteriaceae/genetics Escherichia coli/genetics Frameshift Mutation/genetics Gene Expression/genetics Genetic Complementation Test Molecular Sequence Data Nucleic Acid Conformation Peptide Chain Initiation, Translational/genetics Protein Biosynthesis/genetics Reading Frames/genetics Repetitive Sequences, Nucleic Acid/genetics Shigella dysenteriae/genetics
Chemicals
Codon DNA Transposable Elements
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Polard P
Centre de Recherche en Biochimie et Génétique Cellulaire, CNRS Toulouse, France.
Prère M F
Chandler M
Fayet O
Article Info
Journal
Journal of molecular biology
Abbr.
J Mol Biol
ISSN
0022-2836
Published
1991-12-05
Pages
465-77
Language
English
Region
England
NLM ID
2985088R
Subset
IM
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