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Applied and Environmental Microbiology, July 2009, p. 4419-4426, Vol. 75, No. 13
0099-2240/09/$08.00+0     doi:10.1128/AEM.01523-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Importance of trmE for Growth of the Psychrophile Pseudomonas syringae at Low Temperatures{triangledown} ,{dagger}

Ashish K. Singh, Pavan Kumar Pindi, Smita Dube, V. R. Sundareswaran, and S. Shivaji*

Centre for Cellular and Molecular Biology, Uppal Road, Hyderabad 500 007, India

Received 5 July 2008/ Accepted 25 April 2009

Transposon mutagenesis of Pseudomonas syringae Lz4W, a psychrophilic bacterium capable of growing at temperatures between 2 and 30°C, yielded 30 cold-sensitive mutants, and CSM1, one of these cold-sensitive mutants, was characterized. Growth of CSM1 was retarded when it was cultured at 4°C but not when it was cultured at 22°C and 28°C compared to the growth of wild-type cells, indicating that CSM1 is a cold-sensitive mutant of P. syringae Lz4W. The mutated gene in CSM1 was identified as trmE (coding for tRNA modification GTPase), and evidence is provided that this gene is induced at low temperatures. Further, the cold-inducible nature of the trmE promoter was demonstrated. In addition, the transcription start site and the various regulatory elements of the trmE promoter, such as the –10 region, –35 region, UP element, cold box, and DEAD box, were identified, and the importance of these regulatory elements in promoter activity were confirmed. The importance of trmE in rapid adaptation to growth at low temperatures was further highlighted by plasmid-mediated complementation that alleviated the cold-sensitive phenotype of CSM1.

* Corresponding author. Mailing address: Centre for Cellular and Molecular Biology, Uppal Road, Hyderabad 500 007, India. Phone: 91-40-27192504. Fax: 91-40-27160591. E-mail: shivas{at}ccmb.res.in

{triangledown} Published ahead of print on 8 May 2009.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.

Applied and Environmental Microbiology, July 2009, p. 4419-4426, Vol. 75, No. 13
0099-2240/09/$08.00+0     doi:10.1128/AEM.01523-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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