Mutations in GTP binding protein Obg of Mycoplasma synoviae vaccine strain MS-H: implications in temperature-sensitivity phenotype.
Mycoplasma synoviae strain MS-H, developed by chemical mutagenesis of the Australian field strain 86079/7NS, is a live temperature-sensitive (ts (+)) vaccine used for control of M. synoviae infection in poultry worldwide. Genetic basis of temperature sensitivity and attenuation of MS-H has not been revealed thus far. Comparison of the complete genome sequence of MS-H, its parent strain 86079/7NS and two non-temperature sensitive (ts (-)) reisolates of MS-H revealed a mutation in a highly conserved domain of GTP binding protein Obg of MS-H, with reversion in ts (-) MS-H reisolates. Nucleotide change from G to A at position 369 of the obg gene resulted in an alteration of glycine to arginine at position 123 in Obg fold. Further analysis of the complete obg gene sequence in several MS-H reisolates revealed that a Gly123Arg substitution was associated with alteration in temperature sensitivity phenotype of MS-H. A second mutation, C to T at position 629, in obg gene was found in some of the MS-H reisolates and appeared to suppress the effects of the Gly123Arg substitution. In silico analysis of point mutations revealed that Gly123Arg has highly destabilizing effect on the MS-H Obg structure that can potentially abolish its biological functions in vivo especially at non-permissive temperature. Findings of this study implicate Obg alteration (Gly123Arg) as one of the possible causes of MS-H attenuation/temperature sensitivity and warrant further investigations into exploring the role of Obg-like proteins, an evolutionarily conserved protein from human to bacteria, in the biology of mycoplasmas.
Faculty of Veterinary Science, The University of Melbourne, Werribee, Victoria, Australia.
Muhammad A
MA
Shahid
Philip F
PF
Markham
John F
JF
Markham
Marc S
MS
Marenda
Amir H
AH
Noormohammadi
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D001428:Bacterial Vaccines / Q000235:genetics* / Q000276:immunology; D003062:Codon; D019204:GTP-Binding Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D016680:Genome, Bacterial; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D009175:Mycoplasma Infections / Q000276:immunology / Q000517:prevention & control; D045805:Mycoplasma synoviae / Q000235:genetics* / Q000276:immunology; D010641:Phenotype; D020641:Polymorphism, Single Nucleotide; D011487:Protein Conformation; D055550:Protein Stability; D016415:Sequence Alignment; D013696:Temperature
D001426:Bacterial Proteins; D001428:Bacterial Vaccines; D003062:Codon; C090542:Obg GTP-binding protein, Bacteria; D019204:GTP-Binding Proteins