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Transcriptome changes in Mycoplasma hyopneumoniae during infection
Melissa L Madsen1, Supraja Puttamreddy, Eileen L Thacker
1Department of Veterinary Microbiology and Preventive Medicine, Iowa State University, Ames, IA 50011, USA.
Abstract:
Mycoplasma hyopneumoniae causes swine pneumonia and contributes significantly to the porcine respiratory disease complex. The mechanisms of pathogenesis are difficult to address, since there is a lack of genetic tools, but microarrays are available and can be used to study transcriptional changes that occur during disease as a way to identify important virulence-related genes. Mycoplasmas were collected from bronchial alveolar lavage samples and compared to broth-grown cells using microarrays. Bronchial alveolar lavage was performed on pigs 28 days postinfection, and mycoplasmas were isolated by differential centrifugation. Mycoplasma RNA-enriched preparations were then obtained from total RNA by subtracting eucaryotic ribosomal and messenger RNAs. Labeled cDNAs were generated with mycoplasma open reading frame-specific primers. Nine biological replicates were analyzed. During lung infection, our analysis indicated that 79 M. hyopneumoniae genes were differentially expressed (P < 0.01), at a false-discovery rate of <2.7%. Of the down-regulated genes, 28 of 46 (61%) lacked an assigned function, in comparison to 21 of 33 (63%) of up-regulated genes. Four down-regulated genes and two up-regulated genes encoded putative lipoproteins. secA (mhp295) (P = 0.003) and two glycerol transport permease genes (potA [mhp380; P = 0.006] and ugpA [mhp381; P = 0.003]) were up-regulated in vivo. Elongation factor EF-G (fusA [mhp083]) (P = 0.002), RNA polymerase beta chain (rpoC [mhp635]) (P = 0.003), adenylate kinase (adk [mhp208]) (P = 0.001), prolyl aminoacyl tRNA synthetase (proS [mhp397]) (P = 0.009), and cysteinyl-tRNA synthetase (cysS [mhp661]) (P < 0.001) were down-regulated in vivo.
Insights
This study used microarrays to analyze gene expression in Mycoplasma hyopneumoniae during swine lung infection. Researchers identified 79 differentially expressed genes, offering insights into the pathogen's virulence factors and disease mechanisms.
Area of Science:
- Microbiology
- Animal Science
- Genomics
Background:
- Mycoplasma hyopneumoniae is a key pathogen in swine respiratory disease.
- Understanding its virulence mechanisms is crucial but hindered by a lack of genetic tools.
- Microarray analysis offers a viable method to study transcriptional changes during infection.
Purpose of the Study:
- To identify virulence-related genes of Mycoplasma hyopneumoniae by analyzing transcriptional changes during host infection.
- To compare gene expression profiles of mycoplasmas in vivo (lung infection) versus in vitro (broth culture).
Main Methods:
- Mycoplasma samples were collected from bronchoalveolar lavage fluid of infected pigs.
- Microarray analysis was performed on RNA extracted from mycoplasmas isolated from infected lungs and compared to broth-grown cultures.
- Differential gene expression was analyzed using specific primers, with nine biological replicates.
Main Results:
- A total of 79 Mycoplasma hyopneumoniae genes were found to be differentially expressed (P < 0.01) during lung infection.
- 61% of down-regulated and 63% of up-regulated genes lacked assigned functions, highlighting novel virulence factors.
- Specific genes involved in cell surface components (lipoproteins), glycerol transport (potA, ugpA), and essential cellular processes (secA, fusA, rpoC, adk, proS, cysS) showed significant differential expression.
Conclusions:
- Microarray analysis successfully identified differentially expressed genes in Mycoplasma hyopneumoniae during swine pneumonia.
- The findings provide a foundation for understanding pathogen-host interactions and identifying novel virulence factors.
- This transcriptomic data aids in developing strategies to combat swine respiratory disease complex.
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