A new insertion sequence IS1452 from Acetobacter pasteurianus

Abstract

A new insertion sequence element, IS1452, was found to be associated with inactivation of the alcohol dehydrogenase by insertion in the adhS gene encoding subunit III of the three-component membrane-bound alcohol dehydrogenase complex in Acetobacter pasteurianus. Cloning and sequencing analyses of the mutated subunit III gene locus revealed that IS 1452 was inserted at or near the ribosome-binding sequence of adhS. Analysis of transcription using the chloramphenicol acetyltransferase gene as the reporter indicated that IS1452 abolished transcription of adhS by separating its promoter from the subunit III structural gene. IS1452 was 1411 bp in length and had a terminal inverted repeat of 21 bp. IS1452 contained one long ORF of 416 amino acids rich in basic amino acids. This protein showed homology with a putative transposase, Tra1, of IS701 isolated from the cyanobacterium Calothrix species PCC 7601. Like IS701, IS7452 was found to generate a 4 bp direct repeat at the site of insertion upon transposition. The target site specificity was rather strict, and a CTA(A or G) sequence appeared to be preferentially recognized. Transposition of IS1452 was replicative, since it was accompanied by an increase in the copy number of IS1452. Several strains belonging to the genus Acetobacter also contained IS1452 at varying copy numbers from one to more than ten. These observations suggest that IS1452 is one of the insertion sequences that are responsible for genetic instability leading to deficiencies in various physiological properties in acetic acid bacteria.

Draft Genome Sequence of Acetobacter aceti Strain 1023, a Vinegar Factory Isolate

“…However, A. pasteurianus 386B lacks numerous insertion sequence (IS) elements present in the vinegar strains A. aceti 1023 and A. pasteurianus NBRC 3283 (18). As anticipated from Southern blots (19–21), A. aceti 1023 contains IS 1380 , IS 1452 , and IS 12538 , with minimal copy numbers of 64, 4, and 1, respectively. The adaptation of a common ancestor to different fermentation milieux involved divergent histories of transposable element acquisition in A. aceti 1023 and A. pasteurianus 386B.…”

A New IS 4 Family Insertion Sequence, IS 4Bsu 1, Responsible for Genetic Instability of Poly-γ-Glutamic Acid Production in Bacillus subtilis

“…Some ISs form complex transposable elements, i.e., transposons, by flanking a DNA region encoding antibiotic resistance or containing catabolic or pathogenic genes (3,30,40,44). It is well recognized that transposition of such mobile elements results in insertional mutation or activation of a downstream gene (6,8,13,19,22,23,27,43). As ISs and transposons are often associated with transmissible plasmids and bacteriophages, they distribute in a wide range of bacteria by horizontal transmission.…”

Identification and Distribution of New Insertion Sequences in the Genome of Alkaliphilic Bacillus halodurans C-125

“…Some ISs form composite transposable elements, i.e., transposons, by flanking a DNA region containing antibiotic resistance genes or catabolic or pathogenic genes (4,29,41,48). It is well recognized that transposition of such mobile elements sometimes results in an insertional mutation or activation of a downstream gene (9,12,16,22,25,26,28,47). Since ISs and transposons are often associated with transmissible plasmids and bacteriophages, they have become distributed in a wide range of bacteria by horizontal transmission.…”