DNA Rearrangement of the Shufflon Determines Recipient Specificity in Liquid Mating of IncI1 Plasmid R64

Komano, Teruya; Kim, Su-Ryang; Yoshida, Tetsu; Nisioka, Taizo

doi:10.1006/jmbi.1994.1625

Cited by 48 publications

(61 citation statements)

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“…In Mu and P1 phages, the inversion of the G and C DNA segments controls the alternative expression of S genes for Mu and 19 genes for P1 with different C-terminal segments, which determines the host specificity of both phages (3,14,18). In the shufflon of plasmid R64, the multiple inversion of four DNA segments selects one of seven different pilV genes with different C-terminal segments, which determines recipient specificity in liquid mating of R64 (19,20). In H and type 1 fimbria systems, the inversion of a DNA segment changes the direction of a promoter located within the invertible DNA segment to turn on or turn off the expression of target genes (1,40).…”

Section: Discussionmentioning

confidence: 99%

Integration of bacteriophage Mx8 into the Myxococcus xanthus chromosome causes a structural alteration at the C-terminal region of the IntP protein

et al. 1996

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Mx8 is a generalized transducing phage that infects Myxococcus xanthus cells. This phage is lysogenized inM. xanthus cells by the integration of its DNA into the host chromosome through site-specific recombination. Here, we characterize the mechanism of Mx8 integration into the M. xanthus chromosome. The Mx8 attachment site, attP, the M. xanthus chromosome attachment site, attB, and two phage-host junctions, attL and attR, were cloned and sequenced. Sequence alignments of attP, attB, attL, and attR sites revealed a 29-bp segment that is absolutely conserved in all four sequences. The intP gene of Mx8 was found to encode a basic protein that has 533 amino acids and that carries two domains conserved in site-specific recombinases of the integrase family. Surprisingly, the attP site was located within the coding sequence of the intP gene. Hence, the integration of Mx8 into the M. xanthus chromosome results in the conversion of the intP gene to a new gene designated intR. As a result of this conversion, the 112-residue C-terminal sequence of the intP protein is replaced with a 13-residue sequence. A 3-base deletion within the C-terminal region had no effect on Mx8 integration into the chromosome, while a frameshift mutation with the addition of 1 base at the same site blocked integration activity. This result indicates that the C-terminal region is required for the enzymatic function of the intP product.Myxococcus xanthus is a unique gram-negative bacterium living in soil. M. xanthus cells can undergo multicellular development involving cell-to-cell interactions (for a review, see reference 8). Upon nutritional starvation on a solid surface, cells aggregate to form mounds called fruiting bodies within which rod-shaped cells are converted into spherical or ovoid myxospores.Several bacteriophages that infect M. xanthus cells are known (17). Myxophage Mx8 is a generalized transducing phage of M. xanthus (22). Purified phage particles have 56-kb linear double-stranded DNA molecules with an average terminal redundancy of 4.3 kb (31). Restriction analyses showed that Mx8 phage DNA is circularly permuted (see Fig. 1A). This phage can be lysogenized in M. xanthus cells by integrating its DNA into the host chromosome through site-specific recombination between the attP site on the phage DNA and the attB site on the host chromosome (4, 25). This recombination system has been used to introduce recombinant plasmids into the M. xanthus chromosome, since various plasmids containing a fragment of Mx8 DNA have been shown to stably integrate into the chromosomal attB site (15,29,31). In spite of the effectiveness and wide utilization of the Mx8 attP-mediated integration of plasmids, the integration mechanism is not well understood at present.To reveal the mechanism of Mx8 site-specific recombination in M. xanthus, we analyzed the intP-attP region of Mx8 phage.Comparison of attP, attB, attL, and attR sequences revealed a 29-bp segment that is absolutely conserved. The Mx8 intP gene was shown to encode a basic protein with 533 amino a...

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Section: Discussionmentioning

confidence: 99%

Integration of bacteriophage Mx8 into the Myxococcus xanthus chromosome causes a structural alteration at the C-terminal region of the IntP protein

et al. 1996

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“…pET28b and pET11a (31) were used for overexpression of the pilQ gene. pKK641AЈ (28) carried traABCD and pil genes for R64 thin pilus formation. pKK641AЈ pilQ1 (25) carried the pilQ1 frameshift mutation.…”

Section: Methodsmentioning

confidence: 99%

“…R64 and ColIb-P9 thin pili function in the formation of donor-recipient cell aggregates in liquid matings. Specificity of recipient cells is determined by seven C-terminal segments of PilV adhesins, which are switched through shufflon multiple inversion (27,28). The R64 pilQ product shares amino acid sequence similarity with PulE-VirB11 family NTPases (24).…”

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ATPase Activity and Multimer Formation of PilQ Protein Are Required for Thin Pilus Biogenesis in Plasmid R64

Sakai

Horiuchi

Komano

2001

Journal of Biological Chemistry

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“…The pilV gene product has been putatively identified as a pilus adhesin protein, as in the R64 thin pilus [50]. A shufflon has been observed in both systems, with the Rci recombinase performing a switch between possible alternative C termini of the PilV protein, of which there are two in SPI-7 [51].…”

Section: Type Ivb Pilus Productionmentioning

confidence: 99%

SPI-7: Salmonella’s Vi-Encoding Pathogenicity Island

Seth-Smith

2008

J Infect Dev Ctries

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Salmonella Pathogenicity Island-7 (SPI-7) is a large, mosaic, genetic island, found in several serovars of Salmonella enterica subsp. enterica associated with systemic disease. As well as encoding genes which may aid its own transmission, it carries genes for potential virulence factors such as Vi antigen, SopE effector and type IVB pili. The stability of SPI-7 is of interest with respect to typhoid fever and related vaccines.

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DNA Rearrangement of the Shufflon Determines Recipient Specificity in Liquid Mating of IncI1 Plasmid R64

Cited by 48 publications

References 0 publications

Integration of bacteriophage Mx8 into the Myxococcus xanthus chromosome causes a structural alteration at the C-terminal region of the IntP protein

Integration of bacteriophage Mx8 into the Myxococcus xanthus chromosome causes a structural alteration at the C-terminal region of the IntP protein

ATPase Activity and Multimer Formation of PilQ Protein Are Required for Thin Pilus Biogenesis in Plasmid R64

SPI-7: Salmonella’s Vi-Encoding Pathogenicity Island

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