Isolation and characterization of IS elements repeated in the bacterial chromosome

Matsutani, Sachiko; Ohtsubo, Hideomi; Maeda, Yoshimi; Ohtsubo, Eiichi

doi:10.1016/0022-2836(87)90023-4

Cited by 107 publications

(94 citation statements)

References 36 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Cells harboring cointegrates between the two plasmids were selected in the presence of tetracycline at 420C. The synthetic ISIR was able to mediate cointegration at a frequency of 1.9 x 10-8 per division cycle, which was almost the same frequency reported previously (21). During the chemical synthesis of ISIR, we obtained a mutant (ISIR16) that had an 8-bp deletion at positions 160-167 in the insA coding region (positions 56-328) (see Fig.…”

Section: Methodssupporting

confidence: 70%

See 1 more Smart Citation

Frameshifting is required for production of the transposase encoded by insertion sequence 1.

Sekine

Ohtsubo

1989

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

122

113

View full text Add to dashboard Cite

Insertion sequence IS] has two coding frames, insA and insB, which are essential for its transposition. Here, we show that a frameshifting event in the -1 direction from the 3' end region of the insA frame to an open reading frame (B' frame), extending from the 5' end of the insB frame, is involved in production of the InsA-B'-InsB fusion protein that has IS) transposase activity. The frameshifting event is likely to have occurred at the sequence AAAAAC where the insA frame overlaps the B' frame. Interestingly, this sequence is also present in one of the two sequences identified in retroviruses as frameshift signals for production of transframe polyproteins from the overlapping genes gag-pro or gag-propol.

show abstract

Section: Methodssupporting

confidence: 70%

“…Bacterial strains used were Escherichia coli K12 derivatives, JM109 (20) and JE5519 (13). Plasmid pSAM3, a pUC18 derivative, carried one copy of ISIS derived from the Shigella sonnei chromosome (21). pSEK plasmids were those carrying ISIR mutants.…”

Section: Methodsmentioning

confidence: 99%

Frameshifting is required for production of the transposase encoded by insertion sequence 1.

Sekine

Ohtsubo

1989

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

122

113

View full text Add to dashboard Cite

show abstract

“…This result suggests that the chromosomal copies of IS] in S. dysenteriae may be less active than those of E. coli. Of the three insertions obtained, two were examples of IS600, an element previously isolated by physical means from the Shigella sonnei chromosome (21), while the third (IS911) proved to be an element not described previously.…”

mentioning

confidence: 68%

Transposition in Shigella dysenteriae: isolation and analysis of IS911, a new member of the IS3 group of insertion sequences

1990

View full text Add to dashboard Cite

Twenty-nine clear-plaque mutants of bacteriophage k were isolated from a ShigeUa dysenteriae lysogen. Three were associated with insertions in the cI gene: two were due to insertion of IS600, and the third resulted from insertion of a new element, IS911. IS911 is 1,250 base pairs (bp) long, carries 27-bp imperfect terminal inverted repeats, and generates 3-bp duplications of the target DNA on insertion. It was found in various copy numbers in all four species of Shigella tested and in Escherichia coli K-12 but not in E. coli W. Analysis of IS911-mediated cointegrate molecules indicated that the majority were generated without duplication of IS911.They appeared to result from direct insertion via one end of the element and the neighboring region of DNA, which resembles a terminal inverted repeat of IS911. Nucleotide sequence analysis revealed that IS911 carries two consecutive open reading frames which code for potential proteins showing similarities to those of the IS3 group of elements.In contrast to many strains of Escherichia coli, the genome of members of the closely related genus Shigella carries relatively large numbers of a variety of insertion sequences. It has been estimated, for example, that the four Shigella species carry 40 to 200 copies of insertion sequences closely related to IS] (27). This elevated number of IS-like sequences is not restricted to ISI; it also includes IS2 and IS4 (15; Fayet and Prere, unpublished observations; see reference 11 for review).In order to assess transposition activity from the Shigella dysenteriae genome, we used a simple technique described by Lieb (16) which employs bacteriophage A cI ind as a trap for insertion sequences. By screening for clear-plaque mutants among phages released spontaneously from an E. coli lysogen, it was determined that of 25 mutants, 15 were the result of insertion of IS elements into the cI gene; of these, 7 insertions were of IS], 5 were of IS5, and 3 were of IS3.Since some strains of S. dysenteriae are sensitive to lambdoid phages, we used this technique to obtain some indication of the relative activity of the various IS elements carried by the S. dysenteriae chromosome. We found that the spectrum of cI mutants obtained in S. dysenteriae is very different from that observed in E. coli. No example of an ISl insertion was found, and only 3 of the 29 independent mutants were found to carry insertions in the cI gene. This result suggests that the chromosomal copies of IS] in S. dysenteriae may be less active than those of E. coli. Of the three insertions obtained, two were examples of IS600, an element previously isolated by physical means from the Shigella sonnei chromosome (21), while the third (IS911) proved to be an element not described previously.We present here the nucleotide sequence of IS911 (included in the European Molecular Biology Laboratory sequence data base under accession number X17613), assess its distribution within E. coli K-12, E. coli W, and the four species of Shigella, and analyze some of its transposition properties.

show abstract

“…To ascertain whether any of the hybridization bands observed was due to the presence of IS630, a Shigella IS element that shows homology to one IS200 end (Matsutani et al, 1987), we carried out Southern hybridization analysis using a 5'-32P-labelled 32-mer homologous to the left end of both IS630 and IS200 (Lam & Roth, 1986;Matsutani et al, 1987). Genomic DNAs were digested with AuaI, an endonuclease that does not cut IS200 or IS630 (Casadesiis & Roth, 1989;Matsutani et al, 1987). With this procedure, the number of hybridization bands increased in many S. sonnei and S .…”

Section: Construction and Use Of Is200 Probesmentioning

confidence: 99%

Distribution of insertion sequence IS200 in Salmonella and Shigella

Gibert

Barbé

Casadesús

1990

Journal of General Microbiology

View full text Add to dashboard Cite

Two DNA probes for the detection of insertion sequence IS200 by either Southern blotting or colony hybridization were constructed. One of the probes is a 300 bp EcoRI-Hind111 fragment of IS200 cloned onto pBluescript KS( +); the other is a tail-to-tail dimer of the same fragment cloned onto pUC19. A survey of the presence of IS200 among enteric bacteria revealed that more than 90% of the pathogenic or food-poisoning isolates of Salmonella spp. examined contained one or more copies of insertion sequence IS200, with the exception of the subgenus I Serovar S. agona in which IS200 is not found. Although insertion sequence IS200 was first considered a Salmonellaspecific element, it also exists in many isolates of Shigella sonnei and Shigellajlexneri, but not in Shigella dysenteriae.

show abstract

Isolation and characterization of IS elements repeated in the bacterial chromosome

Cited by 107 publications

References 36 publications

Frameshifting is required for production of the transposase encoded by insertion sequence 1.

Frameshifting is required for production of the transposase encoded by insertion sequence 1.

Transposition in Shigella dysenteriae: isolation and analysis of IS911, a new member of the IS3 group of insertion sequences

Distribution of insertion sequence IS200 in Salmonella and Shigella

Contact Info

Product

Resources

About