Deletion of the prc (tsp) gene provides evidence for additional tail-specific proteolytic activity in Escherichia coli K-12
The Escherichia coli protease Prc (Tsp) exhibits specificity in vitro for proteins with nonpolar carboxyl termini. To determine whether Prc is responsible for the selective degradation in vivo of proteins with nonpolar carboxyl termini, we constructed a prc (tsp) deletion strain. Deletion of the prc gene did not prevent the rapid intracellular degradation of a variant of the amino-terminal domain of λ repressor with a nonpolar carboxyl terminus, even though this protein is a substrate for Prc in vitro. Our results indicate that at least one additional carboxy-terminal-specific proteolytic system must exist in E. coli.
Key wordsSelective protein degradation prc (tsp) deletion strain Carboxyl terminus λ repressor Escherichia coli K-12
Unable to display preview. Download preview PDF.
- Bowie JU, Sauer RT (1989) Identification of C-terminal extensions that protect proteins from intracellular proteolysis. J Biol Chem 264:7596–7602Google Scholar
- Hara H, Yamamoto Y, Higashitani A, Suzuki H, Nishimura Y (1991) Cloning, mapping, and characterization of the Escherichia coli prc gene, which is involved in C-terminal processing of penicillin-binding protein 3. J Bacteriol 173:4799–4813Google Scholar
- Kohara Y, Akiyama K, Isono K (1987) The physical map of the whole E. coli chromosome: application of a new strategy for rapid analysis and sorting of a large genemic library. Cell 50:495–508Google Scholar
- Kornitzer D, Teff D, Altuvia S, Oppenheim AB (1991) Isolation, characterization, and sequence of an Escherichia coli heat shock gene, htpX. J Bacteriol 173:2944–2953Google Scholar
- Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685Google Scholar
- Oden KL, DeVeaux LC, Vibat CRT, Cronan JE Jr., Gennis RB (1990) Genomic replacement in Escherichia coli K-12 using covalently closed circular plasmid DNA. Gene 96:29–36Google Scholar
- Parsell DA, Silber KR, Sauer RT (1990) Carboxy-terminal determinants of intracellular protein degradation. Genes Dev 4:277–286Google Scholar
- Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New YorkGoogle Scholar
- Shurvinton CE, Lloyd RG, Benson FE, Attfield PV (1984) Genetic analysis and molecular cloning of the Escherichia coli ruv gene. Mol Gen Genet 194:322–329Google Scholar
- Silber KR, Keiler KC, Sauer RT (1992) Tsp: a tail-specific protease that selectively degrades proteins with nonpolar C termini. Proc Natl Acad Sci USA 89:295–299Google Scholar
- Singer M, Baker TA, Schnitzler G, Deischel SM, Goel M, Dove W, Jaacks KJ, Grossman AD, Erickson JW, Gross CA (1989) A collection of strains containing genetically linked alternating antibiotic resistance elements for genetic mapping of Escherichia coli. Microbiol Rev 53:1–24Google Scholar