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Journal of Bacteriology, May 2002, p. 2642-2653, Vol. 184, No. 10
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.10.2642-2653.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Functional Analysis of the Signal Recognition Particle in Escherichia coli by Characterization of a Temperature-Sensitive ffh Mutant

Sei-Kyoung Park,1 Fenglei Jiang,2 Ross E. Dalbey,2 and Gregory J. Phillips1*

Department of Microbiology, Iowa State University, Ames, Iowa 50011,1 Department of Chemistry, The Ohio State University, Columbus, Ohio 432102

Received 5 November 2001/ Accepted 15 February 2002

The Ffh protein of Escherichia coli is a 48-kDa polypeptide that is homologous to the SRP54 subunit of the eukaryotic signal recognition particle (SRP). Efforts to understand the function of Ffh in bacteria have depended largely on the use of E. coli strains that allow depletion of the wild-type gene product. As an alternative approach to studying Ffh, a temperature-sensitive ffh mutant was isolated. The ffh-10(Ts) mutation results in two amino acid changes in conserved regions of the Ffh protein, and characterization of the mutant revealed that the cells rapidly lose viability at the nonpermissive temperature of 42°C as well as show reduced growth at the permissive temperature of 30°C. While the ffh mutant is defective in insertion of inner membrane proteins, the export of proteins with cleavable signal sequences is not impaired. The mutant also shows elevated expression of heat shock proteins and accumulates insoluble proteins, especially at 42°C. It was further observed that the temperature sensitivity of the ffh mutant was suppressed by overproduction of 4.5S RNA, the RNA component of the bacterial SRP, by stabilizing the thermolabile protein. Collectively, these results are consistent with a model in which Ffh is required only for localization of proteins integral to the cytoplasmic membrane and suggest new genetic approaches to the study of how the structure of the SRP contributes to its function.

* Corresponding author. Mailing address: Department of Microbiology, 207 Science I Building, Iowa State University, Ames, IA 50011. Phone: (515) 294-1525. Fax: (515) 294-6019. E-mail: gregory{at}iastate.edu.

Journal of Bacteriology, May 2002, p. 2642-2653, Vol. 184, No. 10
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.10.2642-2653.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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