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Genomics and Proteomics

Response of Porphyromonas gingivalis to Heme Limitation in Continuous Culture

Stuart G. Dashper, Ching-Seng Ang, Paul D. Veith, Helen L. Mitchell, Alvin W. H. Lo, Christine A. Seers, Katrina A. Walsh, Nada Slakeski, Dina Chen, J. Patricia Lissel, Catherine A. Butler, Neil M. O'Brien-Simpson, Ian G. Barr, Eric C. Reynolds
Stuart G. Dashper
1Cooperative Research Centre for Oral Health Science, Melbourne Dental School, Bio21 Institute, The University of Melbourne, Victoria, Australia
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Ching-Seng Ang
1Cooperative Research Centre for Oral Health Science, Melbourne Dental School, Bio21 Institute, The University of Melbourne, Victoria, Australia
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Paul D. Veith
1Cooperative Research Centre for Oral Health Science, Melbourne Dental School, Bio21 Institute, The University of Melbourne, Victoria, Australia
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Helen L. Mitchell
1Cooperative Research Centre for Oral Health Science, Melbourne Dental School, Bio21 Institute, The University of Melbourne, Victoria, Australia
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Alvin W. H. Lo
1Cooperative Research Centre for Oral Health Science, Melbourne Dental School, Bio21 Institute, The University of Melbourne, Victoria, Australia
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Christine A. Seers
1Cooperative Research Centre for Oral Health Science, Melbourne Dental School, Bio21 Institute, The University of Melbourne, Victoria, Australia
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Katrina A. Walsh
1Cooperative Research Centre for Oral Health Science, Melbourne Dental School, Bio21 Institute, The University of Melbourne, Victoria, Australia
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Nada Slakeski
1Cooperative Research Centre for Oral Health Science, Melbourne Dental School, Bio21 Institute, The University of Melbourne, Victoria, Australia
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Dina Chen
1Cooperative Research Centre for Oral Health Science, Melbourne Dental School, Bio21 Institute, The University of Melbourne, Victoria, Australia
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J. Patricia Lissel
1Cooperative Research Centre for Oral Health Science, Melbourne Dental School, Bio21 Institute, The University of Melbourne, Victoria, Australia
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Catherine A. Butler
1Cooperative Research Centre for Oral Health Science, Melbourne Dental School, Bio21 Institute, The University of Melbourne, Victoria, Australia
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Neil M. O'Brien-Simpson
1Cooperative Research Centre for Oral Health Science, Melbourne Dental School, Bio21 Institute, The University of Melbourne, Victoria, Australia
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Ian G. Barr
2WHO Collaborating Centre for Reference and Research on Influenza, Parkville, Victoria, Australia
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Eric C. Reynolds
1Cooperative Research Centre for Oral Health Science, Melbourne Dental School, Bio21 Institute, The University of Melbourne, Victoria, Australia
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  • For correspondence: e.reynolds@unimelb.edu.au
DOI: 10.1128/JB.01270-08
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ABSTRACT

Porphyromonas gingivalis is an anaerobic, asaccharolytic, gram-negative bacterium that has essential requirements for both iron and protoporphyrin IX, which it preferentially obtains as heme. A combination of large-scale quantitative proteomic analysis using stable isotope labeling strategies and mass spectrometry, together with transcriptomic analysis using custom-made DNA microarrays, was used to identify changes in P. gingivalis W50 protein and transcript abundances on changing from heme-excess to heme-limited continuous culture. This approach identified 160 genes and 70 proteins that were differentially regulated by heme availability, with broad agreement between the transcriptomic and proteomic data. A change in abundance of the enzymes of the aspartate and glutamate catabolic pathways was observed with heme limitation, which was reflected in organic acid end product levels of the culture fluid. These results demonstrate a shift from an energy-efficient anaerobic respiration to a less efficient process upon heme limitation. Heme limitation also resulted in an increase in abundance of a protein, PG1374, which we have demonstrated, by insertional inactivation, to have a role in epithelial cell invasion. The greater abundance of a number of transcripts/proteins linked to invasion of host cells, the oxidative stress response, iron/heme transport, and virulence of the bacterium indicates that there is a broad response of P. gingivalis to heme availability.

  • Copyright © 2009 American Society for Microbiology
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Response of Porphyromonas gingivalis to Heme Limitation in Continuous Culture
Stuart G. Dashper, Ching-Seng Ang, Paul D. Veith, Helen L. Mitchell, Alvin W. H. Lo, Christine A. Seers, Katrina A. Walsh, Nada Slakeski, Dina Chen, J. Patricia Lissel, Catherine A. Butler, Neil M. O'Brien-Simpson, Ian G. Barr, Eric C. Reynolds
Journal of Bacteriology Jan 2009, 191 (3) 1044-1055; DOI: 10.1128/JB.01270-08

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Response of Porphyromonas gingivalis to Heme Limitation in Continuous Culture
Stuart G. Dashper, Ching-Seng Ang, Paul D. Veith, Helen L. Mitchell, Alvin W. H. Lo, Christine A. Seers, Katrina A. Walsh, Nada Slakeski, Dina Chen, J. Patricia Lissel, Catherine A. Butler, Neil M. O'Brien-Simpson, Ian G. Barr, Eric C. Reynolds
Journal of Bacteriology Jan 2009, 191 (3) 1044-1055; DOI: 10.1128/JB.01270-08
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KEYWORDS

heme
Porphyromonas gingivalis

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