A conserved domain in Escherichia coli Lon protease is involved in substrate discriminator activity.
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.
Ebel
W
W
Department of Microbiology, Oregon State University, Corvallis, Oregon 97331-3804, USA.
Skinner
M M
MM
Dierksen
K P
KP
Scott
J M
JM
Trempy
J E
JE
D049069:ATP-Dependent Proteases; D001426:Bacterial Proteins / Q000378:metabolism; D001483:Base Sequence; D017124:Conserved Sequence; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D029968:Escherichia coli Proteins; D006360:Heat-Shock Proteins / Q000235:genetics* / Q000378:metabolism; D016296:Mutagenesis; D010641:Phenotype; D049070:Protease La; D017422:Sequence Analysis, DNA; D012697:Serine Endopeptidases / Q000235:genetics* / Q000378:metabolism; D013379:Substrate Specificity
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D006360:Heat-Shock Proteins; C053139:sulA protein, E coli; C067572:RcsA protein, E coli; D049069:ATP-Dependent Proteases; D012697:Serine Endopeptidases; C483567:Lon protein, E coli; D049070:Protease La