Cleavage of RseA by RseP requires a carboxyl-terminal hydrophobic amino acid following DegS cleavage

Xiaochun Li, Boyuan Wang, Lihui Feng, Hui Kang, Yang Qi, Jiawei Wang, Yigong Shi

Research output: Contribution to journalArticlepeer-review

35 Scopus citations


Regulated intramembrane proteolysis (RIP) by the Site-2 protease (S2P) results in the release of a transmembrane signaling protein. Curiously, however, S2P cleavage must be preceded by the action of the Site-1 protease (S1P). To decipher the underlying mechanism, we reconstituted sequential, in vitro cleavages of the Escherichia coli transmembrane protein RseA by DegS (S1P) and RseP (S2P). After DegS cleavage, the newly exposed carboxyl-terminal residue Val-148 of RseA plays an essential role for RseP cleavage, and its mutation to charged or dissimilar amino acids crippled the Site-2 cleavage. By contrast, the identity of residues 146 and 147 of RseA has no impact on Site-2 cleavage. These results explain why Site-1 cleavage must precede Site-2 cleavage. Structural analysis reveals that the putative peptide-binding groove in the second, but not the first, PDZ domain of RseP is poised for binding to a single hydrophobic amino acid. These observations suggest that after DegS cleavage, the newly exposed carboxyl terminus of RseA may facilitate Site-2 cleavage through direct interaction with the PDZ domain.

Original languageEnglish (US)
Pages (from-to)14837-14842
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Issue number35
StatePublished - Sep 1 2009


  • Membrane protein
  • PDZ domain
  • Regulated intramembrane proteolysis
  • S2P

ASJC Scopus subject areas

  • General


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