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PMID: 1334529 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Identification of the site of translational frameshifting required for production of the transposase encoded by insertion sequence IS 1.

Molecular & general genetics : MGG ·Vol. 235 ·No. 2-3 ·1992-11-00 ·Pages 317-24

Sekine Y, Nagasawa H, Ohtsubo E

Abstract

Previous genetic analyses indicated that translational frameshifting in the--1 direction occurs within the run of six adenines in the sequence 5'-TTAAAAAACTC-3' at nucleotide positions 305-315 in IS 1, where the two out-of-phase reading frames insA and B'-insB overlap, to produce transposase with a polypeptide segment Leu-Lys-Lys-Leu at residues 84-87. IS 1 mutants with a 1 bp insertion, which encode mutant transposases with an amino acid substitution within the polypeptide segment at residues 84-87, did not efficiently mediate cointegration, except for an IS 1 mutant which encodes a mutant transposase with a Leu-Arg-Lys-Leu segment instead of Leu-Lys-Lys-Leu. An IS 1 mutant with the DNA segment 5'-CTTAAAAACTC-3' at positions 305-315 carrying the termination codon TAA in the B'-insB reading frame could still mediate cointegration, indicating that codon AAA for Lys corresponding to second, third and fourth positions in the run of adenines is the site of frameshifting. The beta-galactosidase activity specified by several IS 1-lacZ fusion plasmids, in which B'-insB is in-frame with lacZ, showed that the region 292-377 is sufficient for frameshifting. The protein produced by frameshifting from the IS 1-lacZ plasmid in fact contained the polypeptide segment Leu-Lys-Lys-Leu encoded by the DNA segment 5'-TTAAAAAACTC-3', indicating that--1 frameshifting does occur within the run of adenines.

Related Genes
lacZ
MeSH Terms
Adenine/analysis Amino Acid Sequence Base Sequence Codon DNA Transposable Elements DNA, Bacterial/genetics Escherichia coli/enzymology,genetics Frameshift Mutation Molecular Sequence Data Nucleotidyltransferases/genetics,metabolism Open Reading Frames Plasmids Protein Biosynthesis Recombinant Fusion Proteins/metabolism Restriction Mapping Transposases beta-Galactosidase/genetics,metabolism
Chemicals
Codon DNA Transposable Elements DNA, Bacterial Recombinant Fusion Proteins Nucleotidyltransferases Transposases beta-Galactosidase Adenine
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Sekine Y
Institute of Applied Microbiology, University of Tokyo, Japan.
Nagasawa H
Ohtsubo E
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33 references, click to expand
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Article Info
Journal
Molecular & general genetics : MGG
Abbr.
Mol Gen Genet
ISSN
0026-8925
DOI
10.1007/BF00279376
Published
1992-11-00
Pages
317-24
Language
English
Region
Germany
NLM ID
0125036
PMCID
PMC7088211
Subset
IM
External Links
PubMed source DOI: 10.1007/BF00279376 PMC Full Text
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