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Infection and Immunity, December 2002, p. 6839-6845, Vol. 70, No. 12
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.12.6839-6845.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

GapA and CrmA Coexpression Is Essential for Mycoplasma gallisepticum Cytadherence and Virulence

L. Papazisi,1,2 S. Frasca Jr.,2 M. Gladd,1,2 X. Liao,1,2 D. Yogev,3 and S. J. Geary1,2*

Center of Excellence for Vaccine Research,1 Department of Pathobiology and Veterinary Science, The University of Connecticut, Storrs, Connecticut 06269,2 Department of Membrane and Ultrastructure Research, The Hebrew University Hadassah Medical School, Jerusalem, Israel3

Received 1 July 2002/ Returned for modification 16 August 2002/ Accepted 18 September 2002

It was previously demonstrated that avirulent Mycoplasma gallisepticum strain Rhigh (passage 164) is lacking three proteins that are expressed in its virulent progenitor, strain Rlow (passage 15). These proteins were identified as the cytadhesin molecule GapA, the putative cytadhesin-related molecule CrmA, and a component of a high-affinity transporter system, HatA. Complementation of Rhigh with wild-type gapA restored expression in the transformant (GT5) but did not restore the cytadherence phenotype and maintained avirulence in chickens. These results suggested that CrmA might play an essential role in the M. gallisepticum cytadherence process. CrmA is encoded by the second gene in the gapA operon and shares significant sequence homology to the ORF6 gene of Mycoplasma pneumoniae, which has been shown to play an accessory role in the cytadherence process. Complementation of Rhigh with wild-type crmA resulted in the transformant (SDCA) that lacked the cytadherence and virulence phenotype comparable to that found in Rhigh and GT5. In contrast, complementation of Rhigh with the entire wild-type gapA operon resulted in the transformant (GCA1) that restored cytadherence to the level found in wild-type Rlow. In vivo pathogenesis trials revealed that GCA1 had regained virulence, causing airsacculitis in chickens. These results demonstrate that both GapA and CrmA are required for M. gallisepticum cytadherence and pathogenesis.

* Corresponding author. Mailing address: Center of Excellence for Vaccine Research and Department of Pathobiology and Veterinary Science, University of Connecticut, 61 N. Eagleville Rd., U-89, Storrs, CT 06269. Phone: (860) 486-0835. Fax: (860) 486-2794. E-mail: geary{at}uconnvm.uconn.edu.

Editor: J. T. Barbieri

Infection and Immunity, December 2002, p. 6839-6845, Vol. 70, No. 12
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.12.6839-6845.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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