TY - JOUR
T1 - Cleavage of RseA by RseP requires a carboxyl-terminal hydrophobic amino acid following DegS cleavage
AU - Li, Xiaochun
AU - Wang, Boyuan
AU - Feng, Lihui
AU - Kang, Hui
AU - Qi, Yang
AU - Wang, Jiawei
AU - Shi, Yigong
N1 - Funding Information:
This work was supported by funds provided by the National Natural Science Foundation of China (81327005, 61361160418, 61575100), the National Foundation of High Technology of China (2012AA020102, 2013AA041201), the National Key Foundation for Exploring Scientific Instruments (2013YQ190467), the Beijing Municipal Natural Science Foundation (4142025), the Beijing Lab Foundation, and the Tsinghua Autonomous Research Foundation (2014Z01001).
PY - 2009/9/1
Y1 - 2009/9/1
N2 - 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.
AB - 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.
KW - Membrane protein
KW - PDZ domain
KW - Regulated intramembrane proteolysis
KW - S2P
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U2 - 10.1073/pnas.0903289106
DO - 10.1073/pnas.0903289106
M3 - Article
C2 - 19706448
AN - SCOPUS:70349299921
SN - 0027-8424
VL - 106
SP - 14837
EP - 14842
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 35
ER -