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Journal of Bacteriology, November 2000, p. 6169-6176, Vol. 182, No. 21
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Nonspecific Adherence by Actinobacillus actinomycetemcomitans Requires Genes Widespread in Bacteria and Archaea

Scott C. Kachlany,1 Paul J. Planet,1 Mrinal K. Bhattacharjee,1 Evyenia Kollia,2 Rob DeSalle,3 Daniel H. Fine,4 and David H. Figurski1,*

Department of Microbiology, College of Physicians and Surgeons, Columbia University, New York, New York 100321; Department of Biology, Bard College, Annandale-on-Hudson, New York 125042; Molecular Laboratories, American Museum of Natural History, New York, New York 100243; and Department of Oral Pathology and Biology, University of Medicine and Dentistry of New Jersey, Newark, New Jersey 071034

Received 12 April 2000/Accepted 5 July 2000

The gram-negative coccobacillus, Actinobacillus actinomycetemcomitans, is the putative agent for localized juvenile periodontitis, a particularly destructive form of periodontal disease in adolescents. This bacterium has also been isolated from a variety of other infections, notably endocarditis. Fresh clinical isolates of A. actinomycetemcomitans form tenacious biofilms, a property likely to be critical for colonization of teeth and other surfaces. Here we report the identification of a locus of seven genes required for nonspecific adherence of A. actinomycetemcomitans to surfaces. The recently developed transposon IS903phi kan was used to isolate mutants of the rough clinical isolate CU1000 that are defective in tight adherence to surfaces (Tad-). Unlike wild-type cells, Tad- mutant cells adhere poorly to surfaces, fail to form large autoaggregates, and lack long, bundled fibrils. Nucleotide sequencing and genetic complementation analysis revealed a 6.7-kb region of the genome with seven adjacent genes (tadABCDEFG) required for tight adherence. The predicted TadA polypeptide is similar to VirB11, an ATPase involved in macromolecular transport. The predicted amino acid sequences of the other Tad polypeptides indicate membrane localization but no obvious functions. We suggest that the tad genes are involved in secretion of factors required for tight adherence of A. actinomycetemcomitans. Remarkably, complete and highly conserved tad gene clusters are present in the genomes of the bubonic plague bacillus Yersinia pestis and the human and animal pathogen Pasteurella multocida. Partial tad loci also occur in strikingly diverse Bacteria and Archaea. Our results show that the tad genes are required for tight adherence of A. actinomycetemcomitans to surfaces and are therefore likely to be essential for colonization and pathogenesis. The occurrence of similar genes in a wide array of microorganisms indicates that they have important functions. We propose that tad-like genes have a significant role in microbial colonization.

* Corresponding author. Mailing address: Department of Microbiology, College of Physicians & Surgeons, Columbia University, 701 W. 168th St., New York, NY 10032. Phone: (212) 305-3425. Fax: (212) 305-1468. E-mail: figurski{at}cuccfa.ccc.columbia.edu.

Journal of Bacteriology, November 2000, p. 6169-6176, Vol. 182, No. 21
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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