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Journal of Bacteriology, December 2006, p. 8178-8188, Vol. 188, No. 23
0021-9193/06/$08.00+0     doi:10.1128/JB.01006-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Global Map of Growth-Regulated Gene Expression in Burkholderia pseudomallei, the Causative Agent of Melioidosis{triangledown}

Fiona Rodrigues,1 Mitali Sarkar-Tyson,2 Sarah V. Harding,2 Siew Hoon Sim,3 Hui Hoon Chua,1 Chi Ho Lin,1 Xu Han,1 R. Krishna M. Karuturi,1 Ken Sung,1 Kun Yu,5 Wei Chen,4 Timothy P. Atkins,2 Richard W. Titball,2 and Patrick Tan1,4*

Genome Institute of Singapore, 60 Biopolis Street, no. 02-01, Genome, Singapore 138672, Republic of Singapore,1 Defence Science and Technology Laboratory, Porton Down, Salisbury, Wiltshire SP4 OJQ, United Kingdom,2 Defence Medical and Environmental Research Institute, DSO National Laboratories, 27 Medical Drive, no. 13-14, Singapore 117510, Republic of Singapore,3 Agenica Research,4 National Cancer Centre, 11 Hospital Drive, Singapore 169610, Republic of Singapore5

Received 9 July 2006/ Accepted 13 September 2006

Many microbial pathogens express specific virulence traits at distinct growth phases. To understand the molecular pathways linking bacterial growth to pathogenicity, we have characterized the growth transcriptome of Burkholderia pseudomallei, the causative agent of melioidosis. Using a fine-scale sampling approach, we found approximately 17% of all B. pseudomallei genes displaying regulated expression during growth in rich medium, occurring as broad waves of functionally coherent gene expression tightly associated with distinct growth phases and transition points. We observed regulation of virulence genes across all growth phases and identified serC as a potentially new virulence factor by virtue of its coexpression with other early-phase virulence genes. serC-disrupted B. pseudomallei strains were serine auxotrophs and in mouse infection assays exhibited a dramatic attenuation of virulence compared to wild-type B. pseudomallei. Immunization of mice with serC-disrupted B. pseudomallei also conferred protection against subsequent challenges with different wild-type B. pseudomallei strains. At a genomic level, early-phase genes were preferentially localized on chromosome 1, while stationary-phase genes were significantly biased towards chromosome 2. We detected a significant level of chromosomally clustered gene expression, allowing us to predict ~100 potential operons in the B. pseudomallei genome. We computationally and experimentally validated these operons by showing that genes in these regions are preferentially transcribed in the same 5'->3' direction, possess significantly shorter intergenic lengths than the overall genome, and are expressed as a common mRNA transcript. The availability of this transcriptome map provides an important resource for understanding the transcriptional architecture of B. pseudomallei.

* Corresponding author. Mailing address: Genome Institute of Singapore, 60 Biopolis Street, no. 02-01, Genome, Singapore 138672, Republic of Singapore. Phone: 65-6-478-8000. Fax: 65-6-478-9003. E-mail: tanbop{at}gis.a-star.edu.sg.

{triangledown} Published ahead of print on 22 September 2006.

Journal of Bacteriology, December 2006, p. 8178-8188, Vol. 188, No. 23
0021-9193/06/$08.00+0     doi:10.1128/JB.01006-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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