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A new insertion sequence IS1452 from Acetobacter pasteurianus

Koichi Kondo1, Sueharu Horinouchi1

  • 1Department of Biotechnology, Division of Agriculture and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113, Japan.

Microbiology (Reading, England)
|February 1, 1997
PubMed
Summary

A novel insertion sequence, IS1452, inactivates alcohol dehydrogenase in Acetobacter pasteurianus by disrupting the adhS gene. This element causes genetic instability and physiological deficiencies in acetic acid bacteria.

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Area of Science:

  • Microbiology
  • Genetics
  • Molecular Biology

Background:

  • Acetic acid bacteria, such as Acetobacter pasteurianus, possess complex metabolic pathways.
  • Alcohol dehydrogenase (ADH) is crucial for their metabolism.
  • Genetic instability can lead to loss of essential functions.

Purpose of the Study:

  • To identify and characterize a novel insertion sequence element, IS1452.
  • To investigate the mechanism by which IS1452 affects gene expression.
  • To understand the role of IS1452 in the genetic instability of acetic acid bacteria.

Main Methods:

  • Cloning and sequencing of the mutated adhS gene locus.
  • Transcriptional analysis using a chloramphenicol acetyltransferase reporter gene.
  • Analysis of IS1452 sequence, including terminal repeats and open reading frames.

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  • Investigation of transposition mechanism and target site specificity.

    • Main Results:

    • IS1452 insertion inactivated the alcohol dehydrogenase by disrupting the adhS gene, likely near the ribosome-binding site.
    • IS1452 abolished adhS transcription by separating the promoter from the structural gene.
    • IS1452 is 1411 bp long with 21 bp terminal inverted repeats and encodes a putative transposase.
    • IS1452 generates 4 bp direct repeats upon transposition, with a preference for CTA(A or G) target sequences.
    • Transposition is replicative, increasing IS1452 copy number.
    • IS1452 is present in multiple strains of Acetobacter.

    Conclusions:

    • IS1452 is a novel insertion sequence element in Acetobacter pasteurianus.
    • IS1452 causes gene inactivation and transcriptional disruption, contributing to genetic instability.
    • This element plays a role in the physiological deficiencies observed in acetic acid bacteria.