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Infection and Immunity, September 2006, p. 5374-5381, Vol. 74, No. 9
0019-9567/06/$08.00+0     doi:10.1128/IAI.00569-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Temperature-Regulated Microcolony Formation by Burkholderia pseudomallei Requires pilA and Enhances Association with Cultured Human Cells

Justin A. Boddey,1,{dagger} Cameron P. Flegg,1 Chris J. Day,2 Ifor R. Beacham,1* and Ian R. Peak1

Institute for Glycomics,1 School of Medical Science, Griffith University, Gold Coast, Queensland, Australia2

Received 6 April 2006/ Returned for modification 2 June 2006/ Accepted 28 June 2006

Burkholderia pseudomallei is the causative agent of melioidosis, a potentially fatal disease that is endemic to Northern Australia and Southeast Asia and is acquired from soil or water. Adherence of B. pseudomallei 08 to cultured cells increases dramatically following prior growth at 30°C or less compared to that following prior growth at 37°C. Here, we show that this occurs almost entirely as the result of microcolony formation (bacterium-bacterium interactions) following growth at 27°C but not at 37°C, which considerably enhances bacterial association with eukaryotic cells. Further, we demonstrate that the type IVA pilin-encoding gene, pilA, is essential for microcolony development by B. pseudomallei 08, and thus optimum association with eukaryotic cells, but is not required for direct adherence (bacterium-cell interactions). In contrast, although the B. pseudomallei genome sequence strain, K96243, also contains transcriptionally active pilA, microcolony formation rarely occurs following growth at either 27°C or 37°C and cell association occurs significantly less than with strain 08. Analysis of pilA transcription in 08 identified that pilA is dramatically upregulated under microcolony-forming conditions, viz., growth at low temperature, and association with eukaryotic cells; the pattern of transcription of pilA in K96243 differed from that in 08. Our study also suggests that biofilm formation by B. pseudomallei 08 and K96243 on polyvinylchloride is not mediated by pilA. Adherence and microcolony formation, and pilA transcription, vary between strains, consistent with known genomic variation in B. pseudomallei, and these phenotypes may be relevant to colonization from the environment.

* Corresponding author. Mailing address: School of Medical Science, Griffith University, Gold Coast Campus, PMB50, Gold Coast Mail Centre, Queensland 9726, Australia. Phone: 61 7 5552 8185. Fax: 011 61 7 5552 8908. E-mail: i.beacham{at}griffith.edu.au.

Editor: V. J. DiRita

{dagger} Present address: The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria, Australia.

Infection and Immunity, September 2006, p. 5374-5381, Vol. 74, No. 9
0019-9567/06/$08.00+0     doi:10.1128/IAI.00569-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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