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PMID:9573050

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Citation

Gottesman, S, Roche, E, Zhou, Y and Sauer, RT (1998) The ClpXP and ClpAP proteases degrade proteins with carboxy-terminal peptide tails added by the SsrA-tagging system. Genes Dev. 12:1338-47

Abstract

Interruption of translation in Escherichia coli can lead to the addition of an 11-residue carboxy-terminal peptide tail to the nascent chain. This modification is mediated by SsrA RNA (also called 10Sa RNA and tmRNA) and marks the tagged polypeptide for proteolysis. Degradation in vivo of lambda repressor amino-terminal domain variants bearing this carboxy-terminal SsrA peptide tag is shown here to depend on the cytoplasmic proteases ClpXP and ClpAP. Degradation in vitro of SsrA-tagged substrates was reproduced with purified components and required a substrate with a wild-type SsrA tail, the presence of both ClpP and either ClpA or ClpX, and ATP. Clp-dependent proteolysis accounts for most degradation of SsrA-tagged amino-domain substrates at 32 degrees C, but additional proteases contribute to the degradation of some of these SsrA-tagged substrates at 39 degrees C. The existence of multiple cytoplasmic proteases that function in SsrA quality-control surveillance suggests that the SsrA tag is designed to serve as a relatively promiscuous signal for proteolysis. Having diverse degradation systems able to recognize this tag may increase degradation capacity, permit degradation of a wide variety of different tagged proteins, or allow SsrA-tagged proteins to be degraded under different growth conditions.

Links

PubMed PMC316764

Keywords

Adenosine Triphosphatases; Amino Acid Sequence; Bacterial Proteins/chemistry; Bacterial Proteins/genetics; Bacterial Proteins/metabolism; Base Sequence; DNA Primers/genetics; Endopeptidase Clp; Escherichia coli/genetics; Escherichia coli/immunology; Escherichia coli/metabolism; Escherichia coli Proteins; Genes, Bacterial; Mutation; Protein Sorting Signals/chemistry; Protein Sorting Signals/genetics; Protein Sorting Signals/metabolism; RNA, Bacterial/metabolism; Recombinant Fusion Proteins/chemistry; Recombinant Fusion Proteins/genetics; Recombinant Fusion Proteins/metabolism; Serine Endopeptidases/genetics; Serine Endopeptidases/metabolism; Signal Transduction; Substrate Specificity; Temperature

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

ECOLI:CLPX

GO:0004176: ATP-dependent peptidase activity

ECO:0000314:

F

Figure 4

complete
CACAO 9069

ECOLI:CLPX

enables

GO:0004176: ATP-dependent peptidase activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

ECOLI:CLPP

GO:0004176: ATP-dependent peptidase activity

ECO:0000314:

F

Figure 4 - ClpP complexes with both ClpA and ClpX to degrade SsrA-tagged proteins in vitro.

complete
CACAO 9066

ECOLI:CLPP

GO:0010498: proteasomal protein catabolic process

ECO:0000315:

P

Figure 5 - Shows ClpP(-) strains degrade the tagged proteins more slowly than wildtype, in vivo.

complete
CACAO 9067

ECOLI:CLPP

enables

GO:0004176: ATP-dependent peptidase activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

ECOLI:CLPP

involved_in

GO:0010498: proteasomal protein catabolic process

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

ECO27:CLPX

GO:0010498: proteasomal protein catabolic process

ECO:0000314:

P

Figure 4 - ClpX works with ClpP in vitro to degrade tagged protein.

complete
CACAO 9059

ECO27:B7UJR0

GO:0010498: proteasomal protein catabolic process

ECO:0000314:

P

Figure 4 - ClpP works with ClpA and ClpX in vitro to degrate SsrA-tagged proteins.

complete
CACAO 9056

ECO27:B7UJR0

GO:1901800: positive regulation of proteasomal protein catabolic process

ECO:0000315:

P

Figure 5 shows ClpP- strains degrade the SsrA-tagged proteins more slowly than wild type strain, in vivo.

complete
CACAO 9057

ECOLI:CLPA

GO:0004176: ATP-dependent peptidase activity

ECO:0000314:

F

Figure 4 - ClpA complexes with ClpP to degrade SsrA-tagged protein, in vitro.

complete
CACAO 9061

ECOLI:CLPA

enables

GO:0004176: ATP-dependent peptidase activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

See also

References

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