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Journal of Bacteriology, April 2004, p. 2393-2401, Vol. 186, No. 8
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.8.2393-2401.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Conserved Target for Group II Intron Insertion in Relaxase Genes of Conjugative Elements of Gram-Positive Bacteria

Jack H. Staddon, Edward M. Bryan,{dagger} Dawn A. Manias, and Gary M. Dunny*

Department of Microbiology, University of Minnesota, Minneapolis, Minnesota 55455

Received 4 August 2003/ Accepted 10 December 2003

The lactococcal group II intron Ll.ltrB interrupts the ltrB relaxase gene within a region that encodes a conserved functional domain. Nucleotides essential for the homing of Ll.ltrB into an intronless version of ltrB are found exclusively at positions required to encode amino acids broadly conserved in a family of relaxase proteins of gram-positive bacteria. Two of these relaxase genes, pcfG from the enterococcal plasmid pCF10 and the ORF4 gene in the streptococcal conjugative transposon Tn5252, were shown to support Ll.ltrB insertion into the conserved motif at precisely the site predicted by sequence homology with ltrB. Insertion occurred through a mechanism indistinguishable from retrohoming. Splicing and retention of conjugative function was demonstrated for pCF10 derivatives containing intron insertions. Ll.ltrB targeting of a conserved motif of a conjugative element suggests a mechanism for group II intron dispersal among bacteria. Additional support for this mechanism comes from sequence analysis of the insertion sites of the E.c.I4 family of bacterial group II introns.

* Corresponding author. Mailing address: Department of Microbiology, University of Minnesota, MMC 196, 420 Delaware St. SE, Minneapolis, MN 55455. Phone: (612) 624-6190. Fax: (612) 626-0623. E-mail: gary-d{at}biosci.cbs.umn.edu.

{dagger} Present address: Virologic, Inc., San Francisco, CA 94030.

Journal of Bacteriology, April 2004, p. 2393-2401, Vol. 186, No. 8
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.8.2393-2401.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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