Formation of SXT Tandem Arrays and SXT-R391 Hybrids

Формирование тандемных массивов SXT и гибридов SXT–R391
Vincent Burrus, Matthew K. Waldor
2004-04-16

RecA-independent recombinationSXT integrative conjugative elementSXT-R391 hybrid elementsconjugative transfertandem SXT arrays
SXT is an integrative and conjugative element (ICE) isolated from Vibrio cholerae. This approximately 100-kb ICE encodes resistance to multiple antibiotics and integrates site specifically into the chromosome. SXT excises from the chromosome to form a circular but nonreplicative extrachromosomal molecule that is required for its transfer. Here we found that a significant fraction of freshly isolated SXT exconjugants contained tandem SXT arrays. There was heterogeneity in the size of the SXT arrays detected in single exconjugant colonies. Some arrays consisted of more than five SXTs arranged in tandem. These extended arrays were unstable and did not persist during serial passages. The mechanism accounting for the generation of SXT arrays is unknown; however, array formation was not dependent upon recA and appeared to depend on conjugative transfer. While such arrays did not alter the transfer frequency of wild-type SXT, they partially complemented the transfer deficiency of a Deltaxis SXT mutant, which is ordinarily unable to generate the extrachromosomal intermediate required for SXT transfer. Exconjugants derived from donor strains that harbored tandem arrays of SXT and R391, an SXT-related element, contained functional hybrid elements that arose from recA-independent recombination between the two ICEs. Thus, arrays of SXT-related elements promote the creation of novel ICEs.
1
Freshly isolated SXT exconjugants frequently contained tandem arrays, with some arrays comprising more than five SXT elements.
2
SXT array sizes varied among individual exconjugant colonies, and extended arrays were unstable during serial passage.
3
SXT tandem-array formation was independent of recA and appeared to require conjugative transfer.
4
Tandem arrays did not change wild-type SXT transfer frequency but partially rescued transfer of a Δxis SXT mutant unable to generate its extrachromosomal intermediate.
5
Tandem arrays of SXT and R391 promoted recA-independent recombination, producing functional hybrid ICEs and novel mobile elements.

SXT integrative and conjugative elements (ICEs), including tandem SXT arrays and SXT-R391 hybrid elements in Vibrio cholerae exconjugants

Formation, stability, conjugative transfer, and recA-independent recombination of tandem SXT-related ICE arrays into novel functional hybrid elements

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2004-04-16
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Vincent Burrus
Matthew K. Waldor
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