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Journal of Bacteriology, December 2002, p. 6929-6941, Vol. 184, No. 24
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.24.6929-6941.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Molecular Genetic Analysis of ICEF, an Integrative Conjugal Element That Is Present as a Repetitive Sequence in the Chromosome of Mycoplasma fermentans PG18

Michael J. Calcutt,{dagger} Michelle S. Lewis,{ddagger} and Kim S. Wise*

Department of Molecular Microbiology and Immunology, University of Missouri—Columbia, Columbia, Missouri 65212

Received 26 June 2002/ Accepted 26 September 2002

Mycoplasma genomes contain compact gene sets that approach the minimal complement necessary for life and reflect multiple evolutionary instances of genomic reduction. Lateral gene transfer may play a critical role in shaping the mobile gene pool in these organisms, yet complex mobile elements have not been reported within this genus. We describe here a large (~23-kb) genetic element with unique features that is present in four copies in the Mycoplasma fermentans PG18 chromosome, accounting for approximately 8% of the genome. These novel elements, designated ICEF (integrative conjugal elements of M. fermentans), resemble conjugative, self-transmissible integrating elements (constins) in that circular, nonreplicative extrachromosomal forms occur in which the left and right termini of the integrated element are juxtaposed and separated by a coupling sequence derived from direct repeats flanking chromosomal copies of ICEF as a result of target site duplication. ICEF contain multiple similarly oriented open reading frames (ORFs), of which some have homology to products of known conjugation genes but others have no known counterparts. Surprisingly, unlike other constins, ICEF lack homologs of known integrases, transposases, or recombinases, suggesting that a novel enzyme may be employed for integration-excision. Skewed distribution and varied sites of chromosomal integration among M. fermentans isolates suggest a role for ICEF in promoting genomic and phenotypic variation in this species. Identification of homologs of terminal ICEF ORFs in two additional mycoplasma species indicates that ICEF is the prototype member of a family of ICE-related elements that may be widespread among pathogenic mycoplasmas infecting diverse vertebrate hosts.

* Corresponding author. Mailing address: Department of Molecular Microbiology and Immunology, School of Medicine, M616 Medical Sciences Building, University of Missouri—Columbia, Columbia, MO 65212. Phone: (573) 882-8138. Fax: (573) 882-4287. E-mail: wisek{at}health.missouri.edu.

{dagger} Present address: Department of Veterinary Pathobiology, University of Missouri—Columbia, Columbia, MO 65211.

{ddagger} Present address: Biology Department, Washington University, St. Louis, MO 63130.

Journal of Bacteriology, December 2002, p. 6929-6941, Vol. 184, No. 24
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.24.6929-6941.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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