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Infection and Immunity, July 2001, p. 4430-4437, Vol. 69, No. 7
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.7.4430-4437.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Cloning, Expression, and Characterization of a Neuraminidase Gene from Arcanobacterium pyogenes

B. H. Jost,* J. G. Songer, and S. J. Billington

Department of Veterinary Science and Microbiology, The University of Arizona, Tucson, Arizona 85721

Received 21 February 2001/Returned for modification 13 April 2001/Accepted 23 April 2001

Arcanobacterium pyogenes is an opportunistic pathogen, associated with suppurative infections in domestic animals. In addition to pyolysin, a pore-forming, cholesterol-binding toxin, A. pyogenes expresses a number of putative virulence factors, including several proteases and neuraminidase activity. A 3,009-bp gene, nanH, was cloned and sequenced and conferred neuraminidase activity on an Escherichia coli host strain. The predicted 107-kDa NanH protein displayed similarity to a number of bacterial neuraminidases and contained the RIP/RLP motif and five copies of the Asp box motif found in all bacterial neuraminidases. Recombinant His-tagged NanH was found to have pH and temperature optima of 5.5 to 6.0 and 55°C, respectively. Insertional deletion of the nanH gene resulted in the reduction, but not absence, of neuraminidase activity, indicating the presence of a second neuraminidase gene in A. pyogenes. NanH was localized to the A. pyogenes cell wall. A. pyogenes adhered to HeLa, CHO, and MDBK cells in a washing-resistant manner. However, the nanH mutant was not defective for adherence to epithelial cells. The role of NanH in host epithelial cell adherence may be masked by the presence of a second neuraminidase in A. pyogenes.

* Corresponding author. Mailing address: Department of Veterinary Science and Microbiology, The University of Arizona, 1117 East Lowell St., Tucson, AZ 85721. Phone: (520) 621 2745. Fax: (520) 621 6366. E-mail: jost{at}u.arizona.edu.

Infection and Immunity, July 2001, p. 4430-4437, Vol. 69, No. 7
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.7.4430-4437.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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