Nucleotide sequence and structural organization of an insertion sequence element (IS231) from Bacillus thuringiensis strain berliner 1715.

(6). Organizational information about the protein has been obtained primarily from genetic approaches, which have defined two regions in IS10 transposase known as Patch I (aa 102-167) and Patch II (aa 243-264) (7). Patch I and Patch II are broadly defined by the positions of mutations that specifically block transposition between the excision and strand transfer (integration) steps, referred to as SOS+Tnsp-(7).Two mutations that confer altered target specificity also occur in these regions, one in Patch I and one in Patch II (8). It has been suggested that these two regions of the protein interact (9).We describe herein a physical investigation of IS10 transposase structure and function. Limited proteolysis shows that transposase protein is organized into two principal domains separated by an exposed linker region that corresponds to a predicted flexible loop. The N-terminal domain (Na,B) is further divisible by more extensive proteolysis into two subdomains (Na and NO3). Southwestern blot analysis demonstrates that the full N-terminal domain Na,B and subdomain NI are both capable of nonspecific DNA binding. Mixtures of purified domains Na,B and C catalyze normal transposition reactions irrespective of the presence of the linker region. If the most N-terminal segment (Na) is missing, however, the only activity detected is the assembly of abnormal synaptic complexes. MATERIALS AND METHODSEnzymatic Digestion of Transposase. IS10 transposase (0.8 mg/ml) was purified as described (10). The purified protein solubilized in buffer A (25 mM Tes, pH 7.5/2 mM EDTA/10 mM dithiothreitol/25 mM Triton X-100/1.5 M NaCl) was treated on ice with trypsin (Boehringer)

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mentioning

confidence: 99%

“…IS10 transposase has 402 aa and shares regions of sequence similarity with other insertion element transposases of the IS4 family (6). Organizational information about the protein has been obtained primarily from genetic approaches, which have defined two regions in IS10 transposase known as Patch I (aa 102-167) and Patch II (aa 243-264) (7).…”

mentioning

confidence: 99%

Structural domains of IS10 transposase and reconstitution of transposition activity from proteolytic fragments lacking an interdomain linker.

Kwon

Chalmers

Kleckner

1995

Proc. Natl. Acad. Sci. U.S.A.

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“…[17], ISSOZ? [18] in E. coli IS231 [19] in Bacillus thuringiensis and ISNZ [20], ISH2 [21] and ISH50 [22] in Halobacterium. We have found no striking similarity in general, but homologies slightly more frequent than expected for random sequence were found in IS5, ISJOR and ISHSU.…”

Section: Nucleotide Sequence Of Is42mentioning

confidence: 99%

IS421, a new insertion sequence in Escherichia coli

1989

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The nucleotide sequence of a new insertion sequence (IS) in Escherichiu co/i, IS421, was determined. It is 1340 bp long and contains inverted repeats of 22 bp at its termini. It is flanked by 13 bp direct repeats apparently generated upon insertion. There are two ORFs longer than 200 bp in IS421. One can encode a polypeptide of 371 amino acids (aa) and the other, which is on the other strand, can encode a polypeptide of 102 aa. The C-terminal part of the 371 aa polypeptide shows some homology to that of transposases encoded in some other known IS elements. The copy number of IS421 in chromosomal DNA was 4 for E. coli K-12 and B, and 5 for E. coli C, as determined by the Southern hybridization of restriction fragments.

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“…pHTA1 contains one copy of IS231 (27) were resistant to Si nuclease treatment and were detected by hybridization with labeled IS231 (Fig. 4B).…”

mentioning

confidence: 99%

“…These sequences have also been detected in other B. thuringiensis strains and have often been found to be associated with crystal protein genes. Mahillon et al (27) sequenced one type of IR from B. thuringiensis berliner which possesses all of the features of an IS structure now named IS231. Furthermore, they showed that this strain of B. thuringiensis contains a family of iso-IS231 elements (26).…”

mentioning

confidence: 99%

A Bacillus thuringiensis subsp. israelensis gene encoding a 125-kilodalton larvicidal polypeptide is associated with inverted repeat sequences

et al. 1988

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A gene encoding a 125-kilodalton (kDa) mosquitocidal delta-endotoxin was cloned from the 72-MDa resident plasmid of Bacillus thuringiensis subsp. israelensis. This gene is similar in its 3' region to the gene encoding the 135-kDa protein previously cloned (C. Bourgouin, A. Klier, and G. Rapoport, Mol. Gen. Genet. 205:390-397, 1986). Escherichia coli recombinant clones harboring the 125-kDa gene were toxic to larvae of the three mosquito species Aedes aegypti, Anopheles stephensi, and Culex pipiens. In addition, the B. thuringiensis subsp. israelensis DNA fragment carrying the 125-kDa protein gene contains two sets of inverted repeat sequences, identified either by the S1 nuclease method or by electron microscopic observation. The structural organization of inverted repeat sequences and of the 125-kDa gene was analyzed and suggests that this B. thuringiensis subsp. israelensis delta-endotoxin gene is located within a transposable element.

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Nucleotide sequence and structural organization of an insertion sequence element (IS231) from Bacillus thuringiensis strain berliner 1715.

Cited by 81 publications

References 42 publications

Structural domains of IS10 transposase and reconstitution of transposition activity from proteolytic fragments lacking an interdomain linker.

Structural domains of IS10 transposase and reconstitution of transposition activity from proteolytic fragments lacking an interdomain linker.

IS421, a new insertion sequence in Escherichia coli

A Bacillus thuringiensis subsp. israelensis gene encoding a 125-kilodalton larvicidal polypeptide is associated with inverted repeat sequences

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