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Journal of Bacteriology, January 2002, p. 111-118, Vol. 184, No. 1
0021-9193/01/$04.00+0 DOI: 10.1128/JB.184.1.111-118.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.
Genetic Screen Yields Mutations in Genes Encoding All Known Components of the Escherichia coli Signal Recognition Particle Pathway
Hongping Tian,
and Jon Beckwith*
Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts 02115
Received 10 July 2001/
Accepted 2 October 2001
We describe the further utilization of a genetic screen that identifies mutations defective in the assembly of proteins into the Escherichia coli cytoplasmic membrane. The screen yielded mutations in each of the known genes encoding components of the E. coli signal recognition particle pathway: ffh, ffs, and ftsY, which encode Ffh, 4.5S RNA, and FtsY, respectively. In addition, the screen yielded mutations in secM, which is involved in regulating levels of the SecA component of the bacteriums protein export pathway. We used a sensitive assay involving biotinylation to show that all of the mutations caused defects in the membrane insertions of three topologically distinct membrane proteins, AcrB, MalF, and FtsQ. Among the mutations that resulted in membrane protein insertion defects, only the secM mutations also showed defects in the translocation of proteins into the E. coli periplasm. Genetic evidence suggests that the S382T alteration of Ffh affects the interaction between Ffh and 4.5S RNA.
* Corresponding author. Mailing address: Department of Microbiology and Genetics, Harvard Medical School, 200 Longwood Ave., Boston, MA 02115. Phone: (617) 432-1920. Fax: (617) 432-7664. E-mail:
jbeckwith{at}hms.harvard.edu.
Present address: HHMI/UCSF Biochemistry & Biophysics, University of California, San Francisco, CA 94143-0448.
Journal of Bacteriology, January 2002, p. 111-118, Vol. 184, No. 1
0021-9193/01/$04.00+0 DOI: 10.1128/JB.184.1.111-118.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.
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