GapA and CrmA coexpression is essential for Mycoplasma gallisepticum cytadherence and virulence

It was previously demonstrated that avirulent Mycoplasma gallisepticum strain R_high (passage 164) is lacking three proteins that are expressed in its virulent progenitor, strain R_low (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 R_high 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 R_high with wild-type crmA resulted in the transformant (SDCA) that lacked the cytadherence and virulence phenotype comparable to that found in R_high and GT5. In contrast, complementation of R_high with the entire wild-type gap_A operon resulted in the transformant (GCA1) that restored cytadherence to the level found in wild-type R_low. 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.