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Journal of Bacteriology, July 2002, p. 3442-3449, Vol. 184, No. 13
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.13.3442-3449.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Natural Transformation and DNA Uptake Signal Sequences in Actinobacillus actinomycetemcomitans

Ying Wang,1 Steve D. Goodman,1 Rosemary J. Redfield,2 and Casey Chen1*

University of Southern California School of Dentistry, Los Angeles, California 90089,1 Department of Zoology, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z42

Received 15 February 2002/ Accepted 10 April 2002

Actinobacillus actinomycetemcomitans is a member of the family Pasteurellaceae and a major causative agent of periodontitis. While several genera from this family are known to be competent for transformation, A. actinomycetemcomitans has yet to be fully characterized. Here we show that the competence of A. actinomycetemcomitans is remarkably similar to that of Haemophilus influenzae. In addition to having a similar frequency of transformation as H. influenzae, A. actinomycetemcomitans competence could also be induced at least 100-fold by cyclic AMP, suggesting that, as in H. influenzae, at least some competence genes are regulated by catabolite repression. Even more intriguing was the discovery of a putative A. actinomycetemcomitans DNA uptake signal sequence (USS) virtually identical to the USS of H. influenzae. Moreover, we provide evidence that this sequence functions in the same capacity as that from H. influenzae; the sequence appears to be required and sufficient for DNA uptake in a variety of assays. Finally, we have taken advantage of this system to develop a simple, highly efficient competence-based method for generating site-directed mutations in the wild-type fimbriated A. actinomycetemcomitans.

* Corresponding author. Mailing address: Division of Primary Oral Health Care, USC School of Dentistry, 925 W. 34th St., Room 4382, Los Angeles, CA 90089-0641. Phone: (213) 740-1075. Fax: (213) 740-6778. E-mail: ccchen{at}hsc.usc.edu.

Journal of Bacteriology, July 2002, p. 3442-3449, Vol. 184, No. 13
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.13.3442-3449.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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