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PMID: 10094703 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

A conserved domain in Escherichia coli Lon protease is involved in substrate discriminator activity.

Journal of bacteriology ·Vol. 181 ·No. 7 ·1999-04-00 ·Pages 2236-43

Ebel W, Skinner MM, Dierksen KP, Scott JM, Trempy JE

Abstract

Lon protease of Escherichia coli regulates a diverse set of physiological responses including cell division, capsule production, plasmid stability, and phage replication. Little is known about the mechanism of substrate recognition by Lon. To examine the interaction of Lon with two of its substrates, RcsA and SulA, we generated point mutations in lon which affected its substrate specificity. The most informative lon mutant overproduced capsular polysaccharide (RcsA stabilized) yet was resistant to DNA-damaging agents (SulA degraded). Immunoblots revealed that RcsA protein persisted in this mutant whereas SulA protein was rapidly degraded. The mutant contains a single-base change within lon leading to a single amino acid change of glutamate 240 to lysine. E240 is conserved among all Lon isolates and resides in a charged domain that has a high probability of adopting a coiled-coil conformation. This conformation, implicated in mediating protein-protein interactions, appears to confer substrate discriminator activity on Lon. We propose a model suggesting that this coiled-coil domain represents the discriminator site of Lon.

MeSH Terms
ATP-Dependent Proteases Bacterial Proteins/metabolism Base Sequence Conserved Sequence Escherichia coli/enzymology,genetics Escherichia coli Proteins Heat-Shock Proteins/genetics,metabolism Mutagenesis Phenotype Protease La Sequence Analysis, DNA Serine Endopeptidases/genetics,metabolism Substrate Specificity
Chemicals
Bacterial Proteins Escherichia coli Proteins Heat-Shock Proteins sulA protein, E coli RcsA protein, E coli ATP-Dependent Proteases Serine Endopeptidases Lon protein, E coli Protease La
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Ebel W
Department of Microbiology, Oregon State University, Corvallis, Oregon 97331-3804, USA.
Skinner M M
Dierksen K P
Scott J M
Trempy J E
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/JB.181.7.2236-2243.1999
Published
1999-04-00
Pages
2236-43
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC93638
Subset
IM
External Links
PubMed source DOI: 10.1128/JB.181.7.2236-2243.1999 PMC Full Text View journal details
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