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Tn5386, a Novel Tn916-Like Mobile Element in Enterococcus faecium D344R That Interacts with Tn916 To Yield a Large Genomic Deletion

Medical and Research Services, Louis Stokes Cleveland Department of Veterans Affairs Medical Center,¹ Department of Medicine, Case Western Reserve University, Cleveland, Ohio²

Received 18 February 2005/ Accepted 25 April 2005

We describe Tn5386, a novel ca.-29-kb Tn916-like mobile element discovered to occur in ampicillin-resistant, Tn916-containing Enterococcus faecium D344R. PCR amplification experiments after overnight growth with or without tetracycline revealed "joint" regions of circularized Tn5386 composed of 6-bp sequences linking different transposon termini. In one case (no tetracycline), the termini were consistent with those derived by target site analysis of the integrated element. In the other case, the termini were virtually identical in distance from the integrase binding regions, as seen with Tn916. These data are consistent with a model in which one PCR product results from the action of Tn5386 integrase, whereas the other results from the action of the Tn916 integrase on Tn5386. Spontaneous conversion of D344R to an ampicillin-susceptible phenotype (D344SRF) was associated with a 178-kb deletion extending from the left end of Tn5386 to the left end of Tn916. Examination of the Tn5386 junction after the large deletion event suggests that the deletion resulted from an interaction between the nonintegrase ends of Tn5386 and Tn916. The terminus of Tn5386 identified in this reaction suggested that it may have resulted from the activity of the Tn916 integrase (Int_Tn916). The "joint" of the circular element resulting from this excision was amplifiable from D344R, the sequence of which revealed a heteroduplex consistent with Int_Tn916-mediated excision. In contrast, Tn5386 joints amplified from ampicillin-susceptible D344SRF revealed ends consistent with Tn5386 integrase activity, reflecting the absence of Tn916 from this strain. Tn5386 represents a new member of the Tn916 transposon family. Our data suggest that excision of Tn5386 can be catalyzed by the Tn916 integrase and that large genomic deletions may result from the interaction between these heterologous elements.

This article has been cited by other articles:

• Rice, L. B., Carias, L. L., Hutton-Thomas, R., Rudin, S. (2007). Interaction of Related Tn916-Like Transposons: Analysis of Excision Events Promoted by Tn916 and Tn5386 Integrases. J. Bacteriol. 189: 3909-3917 [Abstract] [Full Text]  
• Rice, L. B., Carias, L. L., Rudin, S., Lakticova, V., Wood, A., Hutton-Thomas, R. (2005). Enterococcus faecium Low-Affinity pbp5 Is a Transferable Determinant. Antimicrob. Agents Chemother. 49: 5007-5012 [Abstract] [Full Text]