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Genetics and Molecular Biology |
School of Health Science, Griffith University, PMB 50 Gold Coast Mail Centre, Gold Coast, Qld 4217, Australia1
Author for correspondence: Ifor R. Beacham. Tel: +61 7 5594 8087. Fax: +61 7 5594 8908. e-mail: i.beacham{at}mailbox.gu.edu.au
The production of extracellular enzymes by Pseudomonas fluorescens is important with respect to phytopathogenesis and, in the case of psychrotrophic strains, food spoilage. The production of extracellular protease has been previously reported to be dependent on temperature in psychrotrophic strains of P. fluorescens; production is decreased above the optimum growth temperature with a relatively small change in growth rate. In this work, a transposon mutant of P. fluorescens LS107d2 has been isolated which, in contrast to the wild-type strain, is completely protease deficient at 29 °C, above the optimum growth temperature of 25 °C, but which produces protease at 23 °C. Further analysis revealed that this mutation is in a gene (prtR) which is part of a dicistronic operon, prtIR, in which the two genes are translationally coupled. Evidence is presented that prtI encodes a sigma factor related to others involved in extracytoplasmic functions (ECF sigma factors) and that prtR encodes a novel transmembrane activator of PrtI. PrtI, like PrtR, is also required for protease production at 29 °C but not at 23 °C. Analysis of the amino acid sequence of PrtR indicates that it is functionally related to a group of membrane-associated anti-sigma factors and a few transmembrane regulators, but is not significantly sequence related. Complementation analysis indicates that PrtR may also interact with sigma factors other than PrtI. The promoter region of the protease-encoding gene (aprX) in LS107d2 has been identified and has sequence features which could indicate interaction with either an ECF sigma factor or a primary sigma factor.
Keywords: protease, Pseudomonas fluorescens, ECF sigma factor, transmembrane activator, temperature regulation
The GenBank accession numbers for the sequences reported in this paper are AF228766 and AF228767.
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