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Organization of the Mycoplasma synoviae WVU 1853T vlhA gene locus
Nathan Jeffery1, Glenn F Browning, Amir H Noormohammadi
1CSIRO Livestock Indstries, F.D. McMaster Laboratory Chiswick, Armidale, NSW 2350, Australia.
Abstract:
Mycoplasma synoviae cause respiratory disease and synovitis in poultry. It has two major membrane antigens of approximately 45 to 50 kDa, MSPA and MSPB. Both MSPA and MSPB ar encoded by a single gene, vlhA (variable lipoprotein and haemagglutinin), possibly with a post-translational cleavage generating the two proteins. The vlhA gene belongs to a large multigene family, but only one vlhA gene is expressed in any single cell; the other vlhA genes/fragments are transcriptionally silent. In order to characterize the vlhA gene locus, DNA fragments were cloned from three different M. synoviae WVU 1853(T) genomic DNA libraries and their nucleotide sequences were fully or partially determined. Analysis of the resultant nucleotide sequences identified a transcriptional termination signal for the expressed vlhA gene, determined the genes located downstream of the expressed vlhA gene and revealed that vlhA pseudogenes were arranged as tandem repeats upstream of the expressed vlhA gene. In order to determine whether vlhA was expressed as a monocistronic or polycistronic message, RNA from two M. synoviae clones expressing truncated or full-length versions of the vlhA gene product were purified and examined by northern blotting. Messages corresponding to the full length of the vlhA gene (approximately 2.4 kb) were detected in both clones, suggesting that truncation of the vlhA gene product was probably post-transcriptional. These studies have revealed the organization of the M. synoviae vlhA gene locus and established that the vlhA gene transcript is monocistronic.
Insights
Mycoplasma synoviae expresses the vlhA gene, encoding major membrane antigens MSPA and MSPB. This study characterized the vlhA gene locus, revealing its organization and monocistronic transcript, crucial for understanding poultry respiratory disease.
Area of Science:
- Veterinary Microbiology
- Molecular Biology
- Genomics
Background:
- Mycoplasma synoviae is a significant poultry pathogen causing respiratory disease and synovitis.
- The bacterium possesses major membrane antigens MSPA and MSPB, encoded by the vlhA gene.
- The vlhA gene is part of a multigene family with only one expressed copy per cell.
Purpose of the Study:
- To characterize the genomic locus of the expressed vlhA gene in Mycoplasma synoviae.
- To elucidate the transcriptional organization of the vlhA gene.
- To investigate the expression of vlhA and its encoded antigens.
Main Methods:
- Genomic DNA library construction and nucleotide sequencing of M. synoviae WVU 1853(T).
- Bioinformatic analysis to identify gene structures, regulatory elements, and pseudogenes.
- Northern blotting to analyze vlhA gene transcript size and expression.
Main Results:
- The expressed vlhA gene locus organization was determined, including upstream tandem repeat pseudogenes and downstream genes.
- A transcriptional termination signal for the expressed vlhA gene was identified.
- Northern blot analysis confirmed a monocistronic, full-length (approx. 2.4 kb) vlhA transcript, indicating post-transcriptional modification of the protein product.
Conclusions:
- The M. synoviae vlhA gene locus is organized with pseudogenes and downstream genes.
- The vlhA gene is transcribed as a monocistronic message.
- Understanding the vlhA gene locus and expression is vital for developing strategies against Mycoplasma synoviae infections in poultry.
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