title: Cleavage of RseA by RseP requires a carboxyl-terminal hydrophobic amino acid following DegS cleavage.
abstract: 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.
authors.affiliation: Ministry of Education Protein Science Laboratory and Center for Structural Biology, Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing 100084, China.
authors.forename: Xiaochun
authors.initials: X
authors.lastname: Li
authors.forename: Boyuan
authors.initials: B
authors.lastname: Wang
authors.forename: Lihui
authors.initials: L
authors.lastname: Feng
authors.forename: Hui
authors.initials: H
authors.lastname: Kang
authors.forename: Yang
authors.initials: Y
authors.lastname: Qi
authors.forename: Jiawei
authors.initials: J
authors.lastname: Wang
authors.forename: Yigong
authors.initials: Y
authors.lastname: Shi
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D053829:Amyloid Precursor Protein Secretases / Q000378:metabolism; D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D017124:Conserved Sequence; D010450:Endopeptidases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D011485:Protein Binding; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary; D016415:Sequence Alignment; D013379:Substrate Specificity; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*
chemical_list: D000596:Amino Acids; D029968:Escherichia coli Proteins; D008565:Membrane Proteins; C433802:RseA protein, E coli; D014157:Transcription Factors; C489349:degS protein, Escherichia coli; D053829:Amyloid Precursor Protein Secretases; D010450:Endopeptidases; C440001:RseP protein, E coli
references.citation: Curr Opin Struct Biol. 2008 Aug;18(4):432-41
references.pmid: 18440799
references.citation: Essays Biochem. 2002;38:155-68
references.pmid: 12463168
references.citation: Mol Cell. 1998 Oct;2(4):505-14
references.pmid: 9809072
references.citation: Proc Natl Acad Sci U S A. 2007 Mar 6;104(10):3771-6
references.pmid: 17360428
references.citation: Cell. 1997 May 2;89(3):331-40
references.pmid: 9150132
references.citation: Mol Microbiol. 2004 Apr;52(2):427-35
references.pmid: 15066031
references.citation: Annu Rev Genet. 2004;38:709-24
references.pmid: 15568990
references.citation: J Biol Chem. 1997 May 9;272(19):12778-85
references.pmid: 9139737
references.citation: Cell. 2003 Apr 4;113(1):61-71
references.pmid: 12679035
references.citation: Genes Dev. 2002 Aug 15;16(16):2147-55
references.pmid: 12183368
references.citation: Cell. 2004 May 14;117(4):483-94
references.pmid: 15137941
references.citation: Genes Dev. 2007 Oct 15;21(20):2659-70
references.pmid: 17938245
references.citation: Cell. 1996 Jun 28;85(7):1037-46
references.pmid: 8674110
references.citation: Cell. 2007 Nov 2;131(3):572-83
references.pmid: 17981123
references.citation: Science. 2004 Aug 20;305(5687):1119-23
references.pmid: 15326347
references.citation: J Biol Chem. 1998 Jul 10;273(28):17801-9
references.pmid: 9651382
references.citation: Cell. 2000 Feb 18;100(4):391-8
references.pmid: 10693756
references.citation: EMBO J. 2004 Nov 10;23(22):4434-42
references.pmid: 15496982
references.citation: Microbes Infect. 2006 Jun;8(7):1882-8
references.pmid: 16731018
references.citation: EMBO J. 2003 Dec 1;22(23):6389-98
references.pmid: 14633997
references.citation: Genes Dev. 2002 Aug 15;16(16):2156-68
references.pmid: 12183369
references.citation: J Biol Chem. 2008 Dec 12;283(50):35042-52
references.pmid: 18945679
references.citation: Neuron. 2002 May 16;34(4):499-502
references.pmid: 12062033
references.citation: Mol Microbiol. 2004 May;52(3):613-9
references.pmid: 15101969
references.citation: J Cell Sci. 2001 Sep;114(Pt 18):3219-31
references.pmid: 11591811
references.citation: J Biol Chem. 2008 Apr 11;283(15):9562-70
references.pmid: 18268014
references.citation: Curr Opin Genet Dev. 2002 Oct;12(5):512-8
references.pmid: 12200155
references.citation: Protein Sci. 2006 Jan;15(1):84-93
references.pmid: 16322567
references.citation: Cell. 2005 Aug 12;122(3):435-47
references.pmid: 16096062
references.citation: Mol Cell. 1997 Dec;1(1):47-57
references.pmid: 9659902