title: A novel mechanism for control of antigenic variation in the haemagglutinin gene family of mycoplasma synoviae.
abstract: High-frequency phase and antigenic variation of homologous lipoprotein haemagglutinins has been seen in both the major avian mycoplasma pathogens, Mycoplasma synoviae and Mycoplasma gallisepticum. The expression and, hence, antigenic variation of the pMGA gene family (encoding these lipoproteins in M. gallisepticum) is controlled by variation in the length of a trinucleotide repeat motif 5' to the promoter of each gene. However, such a mechanism was not detected in preliminary observations on M. synoviae. Thus, the basis for control of variation in the vlhA gene family (which encodes the homologous haemagglutinin in M. synoviae) was investigated to enable comparison with its homologue in M. gallisepticum and with other lipoprotein gene families in mycoplasmas. The start point of transcription was identified 119 bp upstream of the initiation codon, but features associated with control of transcription in other mycoplasma lipoprotein genes were not seen. Comparison of three copies of vlhA revealed considerable sequence divergence at the 3' end of the gene, but conservation of the 5' end. Southern blot analysis of M. synoviae genomic DNA revealed that the promoter region and part of the conserved 5' coding sequence occurred as a single copy, whereas the remainder of the coding sequence occurred as multiple copies. A 9.7 kb fragment of the genome was found to contain eight tandemly repeated regions partially homologous to vlhA, all lacking the putative promoter region and the single-copy 5' end of vlhA, but extending over one of four distinct overlapping regions of the 3' coding sequence. Examination of sequential clones of M. synoviae established that unidirectional recombination occurs between the pseudogenes and the expressed vlhA, with duplication of pseudogene sequence and loss of the corresponding region previously seen in the expressed gene. Expression of the 5' end of two variants of the vlhA gene showed that they differed in their reaction with monoclonal antibodies specific for this region. These data suggest that the control of vlhA antigenic variation in M. synoviae is achieved by multiple gene conversion events using a repertoire of coding sequences to generate a chimeric expressed gene, with the greatest potential for variation generated in the region encoding the haemagglutinin. Thus, completely distinct mechanisms have been adopted to control antigenic variation in homologous gene families.
authors.affiliation: School of Veterinary Science, The University of Melbourne, Cnr. Flemington Road and Park Drive, Parkville, Victoria, Australia. amirh@clyde.its.unimelb.edu.au
authors.forename: A H
authors.initials: AH
authors.lastname: Noormohammadi
authors.forename: P F
authors.initials: PF
authors.lastname: Markham
authors.forename: A
authors.initials: A
authors.lastname: Kanci
authors.forename: K G
authors.initials: KG
authors.lastname: Whithear
authors.forename: G F
authors.initials: GF
authors.lastname: Browning
mesh_terms: D000595:Amino Acid Sequence; D000940:Antigenic Variation; D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D017124:Conserved Sequence; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D014644:Genetic Variation; D006388:Hemagglutinins / Q000235:genetics*; D037102:Lectins; D008969:Molecular Sequence Data; D005810:Multigene Family; D009174:Mycoplasma / Q000235:genetics*; D016366:Open Reading Frames; D011401:Promoter Regions, Genetic; D011544:Pseudogenes / Q000235:genetics; D011995:Recombination, Genetic; D012091:Repetitive Sequences, Nucleic Acid; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; D006388:Hemagglutinins; D037102:Lectins; C079720:M9 protein, Mycoplasma gallisepticum; C406455:VlhA protein, Mycoplasma synoviae