Insertion sequence IS2 associated with int-constitutive mutants of bacteriophage lambda

Pilacinski, William P.; Mosharrafa, E.; Edmundson, R.; Zissler, J.; Fiandt, M.; Szybalski, Waclaw

doi:10.1016/0378-1119(77)90073-7

Cited by 51 publications

(14 citation statements)

References 18 publications

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“…Multiple insertions into the promoters of the galOPETK operon (1,27,32), the divergent argECBH operon (15), the xis and int genes of bacteriophage lambda (25,30), and the ceL4BCDF operon (29) and single insertions into the upstream regulatory regions of the lac operon (9), ampC (18), ilvA of Pseudomonas cepacia (2), the Tetr gene in pBR322 (6), and his3 and trp5 of Saccharomyces cerevisiae (4,5) have been reported. In these instances, IS2 exhibits either a strong polar effect (19) in orientation I (32) or an ability to promote transcription in adjacent genes when inserted in orientation IL (15,32; see also references 13 and 16 for comprehensive reviews).…”

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confidence: 99%

Transposition of IS2 into the hemB gene of Escherichia coli K-12

Lewis

Persaud

et al. 1994

J Bacteriol

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Genetic studies of the hemB gene in Escherichia coli have resulted in the recovery of both stable and unstable mutant strains. The stable strains have been shown to result from large deletions. This study demonstrates that unstable strains result from the insertion of transposable element IS2 primarily into the 5' region of the structural gene; the instability results from precise excision of the element, producing strains with both high and low frequencies of reversion. This first report of IS2 insertion into hemB suggests that this gene may be a preferred target for insertion of this transposable element.This study describes the first report of the transposition of IS2 into the hemB gene of Escherichia coli. Early genetic studies on the characterization and mapping of hemB described the recovery of many deletion mutations associated with this gene (3,23,36). In recent studies of microcolony, respiratory-deficient (Res-), gentamicin-resistant mutants of E. coli, hemB was shown at the molecular level to be a hot spot for spontaneous mutations. More than 80% of 150 independently isolated Res -mutants resulted from mutations which mapped to hemB, and a molecular analysis of 33 of these revealed that 73% of them resulted from both deletions and insertions (20). Further analysis of the insertion mutations is reported here.The transposable element IS2 is a 1.3-kb insertion sequence (14, 31) which occurs naturally in E. coli K-12 and is best known for its ability to generate mutations by insertion into the regulatory regions of genes. Multiple insertions into the promoters of the galOPETK operon (1, 27, 32), the divergent argECBH operon (15), the xis and int genes of bacteriophage lambda (25,30), and the ceL4BCDF operon (29) and single insertions into the upstream regulatory regions of the lac operon (9), ampC (18), ilvA of Pseudomonas cepacia (2), the Tetr gene in pBR322 (6), and his3 and trp5 of Saccharomyces cerevisiae (4, 5) have been reported. In these instances, IS2 exhibits either a strong polar effect (19) in orientation I (32) or an ability to promote transcription in adjacent genes when inserted in orientation IL (15, 32; see also references 13 and 16 for comprehensive reviews). Less often reported is the insertion of IS2 into the coding regions of genes, e.g., argB (7), galK (39), and reLA (24), in which it disrupts the coding sequences. Despite extensive studies of IS2 insertions into the phage X (34) and phage P1 (38-40) chromosomes, a molecular basis for the nonrandom pattern of its insertional specificity is still unclear.We show here, using restriction digestion, Southern hybridization, and PCR amplification followed by the direct sequencing of the PCR products that (i) all of the insertion mutations studied for hemB resulted from the transposition of IS2 into the structural gene; (ii) six of the eight insertion events studied * Corresponding

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Transposition of IS2 into the hemB gene of Escherichia coli K-12

Lewis

Persaud

et al. 1994

J Bacteriol

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“…These elements, comprised of a transposase gene and inverted repeats, create flanking direct repeats following insertion [68]. Many mutants of well-characterized phages have been found to carry ISs, including λ and Mu [69,70]. However, it is relatively rare for wildtype phages to carry ISs because they can interfere with gene expression [71].…”

Section: Resultsmentioning

confidence: 99%

Genomic analysis and relatedness of P2-like phages of the Burkholderia cepacia complex

2010

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BackgroundThe Burkholderia cepacia complex (BCC) is comprised of at least seventeen Gram-negative species that cause infections in cystic fibrosis patients. Because BCC bacteria are broadly antibiotic resistant, phage therapy is currently being investigated as a possible alternative treatment for these infections. The purpose of our study was to sequence and characterize three novel BCC-specific phages: KS5 (vB_BceM-KS5 or vB_BmuZ-ATCC 17616), KS14 (vB_BceM-KS14) and KL3 (vB_BamM-KL3 or vB_BceZ-CEP511).ResultsKS5, KS14 and KL3 are myoviruses with the A1 morphotype. The genomes of these phages are between 32317 and 40555 base pairs in length and are predicted to encode between 44 and 52 proteins. These phages have over 50% of their proteins in common with enterobacteria phage P2 and so can be classified as members of the Peduovirinae subfamily and the "P2-like viruses" genus. The BCC phage proteins similar to those encoded by P2 are predominantly structural components involved in virion morphogenesis. As prophages, KS5 and KL3 integrate into an AMP nucleosidase gene and a threonine tRNA gene, respectively. Unlike other P2-like viruses, the KS14 prophage is maintained as a plasmid. The P2 E+E' translational frameshift site is conserved among these three phages and so they are predicted to use frameshifting for expression of two of their tail proteins. The lysBC genes of KS14 and KL3 are similar to those of P2, but in KS5 the organization of these genes suggests that they may have been acquired via horizontal transfer from a phage similar to λ. KS5 contains two sequence elements that are unique among these three phages: an ISBmu2-like insertion sequence and a reverse transcriptase gene. KL3 encodes an EcoRII-C endonuclease/methylase pair and Vsr endonuclease that are predicted to function during the lytic cycle to cleave non-self DNA, protect the phage genome and repair methylation-induced mutations.ConclusionsKS5, KS14 and KL3 are the first BCC-specific phages to be identified as P2-like. As KS14 has previously been shown to be active against Burkholderia cenocepacia in vivo, genomic characterization of these phages is a crucial first step in the development of these and similar phages for clinical use against the BCC.

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“…In contrast, some of the earliest experiments on mobile elements demonstrated their ability to relocate and generate novel transcriptional signals in prokaryotes and eukaryotes 219 - 221 …”

Section: Beyond Horizontal Transfer: Intracellular Natural Genetic Enmentioning

confidence: 96%

Constraint and opportunity in genome innovation

Shapiro

2013

RNA Biology

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The development of rigorous molecular taxonomy pioneered by Carl Woese has freed evolution science to explore numerous cellular activities that lead to genome change in evolution. These activities include symbiogenesis, inter- and intracellular horizontal DNA transfer, incorporation of DNA from infectious agents, and natural genetic engineering, especially the activity of mobile elements. This article reviews documented examples of all these processes and proposes experiments to extend our understanding of cell-mediated genome change.

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Insertion sequence IS2 associated with int-constitutive mutants of bacteriophage lambda

Cited by 51 publications

References 18 publications

Transposition of IS2 into the hemB gene of Escherichia coli K-12

Transposition of IS2 into the hemB gene of Escherichia coli K-12

Genomic analysis and relatedness of P2-like phages of the Burkholderia cepacia complex

Constraint and opportunity in genome innovation

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