title: Deciphering the assembly of the Yersinia type III secretion injectisome.
abstract: The assembly of the Yersinia enterocolitica type III secretion injectisome was investigated by grafting fluorescent proteins onto several components, YscC (outer-membrane (OM) ring), YscD (forms the inner-membrane (IM) ring together with YscJ), YscN (ATPase), and YscQ (putative C ring). The recombinant injectisomes were functional and appeared as fluorescent spots at the cell periphery. Epistasis experiments with the hybrid alleles in an array of injectisome mutants revealed a novel outside-in assembly order: whereas YscC formed spots in the absence of any other structural protein, formation of YscD foci required YscC, but not YscJ. We therefore propose that the assembly starts with YscC and proceeds through the connector YscD to YscJ, which was further corroborated by co-immunoprecipitation experiments. Completion of the membrane rings allowed the subsequent assembly of cytosolic components. YscN and YscQ attached synchronously, requiring each other, the interacting proteins YscK and YscL, but no further injectisome component for their assembly. These results show that assembly is initiated by the formation of the OM ring and progresses inwards to the IM ring and, finally, to a large cytosolic complex.
authors.affiliation: Infection Biology, Biozentrum der Universität Basel, Basel, Switzerland.
authors.forename: Andreas
authors.initials: A
authors.lastname: Diepold
authors.forename: Marlise
authors.initials: M
authors.lastname: Amstutz
authors.forename: Sören
authors.initials: S
authors.lastname: Abel
authors.forename: Isabel
authors.initials: I
authors.lastname: Sorg
authors.forename: Urs
authors.initials: U
authors.lastname: Jenal
authors.forename: Guy R
authors.initials: GR
authors.lastname: Cornelis
mesh_terms: D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism*; D047468:Immunoprecipitation; D015008:Yersinia enterocolitica / Q000235:genetics / Q000378:metabolism*
chemical_list: D000251:Adenosine Triphosphatases
references.citation: Nat Rev Microbiol. 2006 Nov;4(11):811-25
references.pmid: 17041629
references.citation: Science. 2004 Nov 5;306(5698):1040-2
references.pmid: 15528446
references.citation: Mol Microbiol. 2000 Sep;37(6):1494-503
references.pmid: 10998179
references.citation: Microbes Infect. 2002 Jan;4(1):75-82
references.pmid: 11825778
references.citation: J Bacteriol. 2001 Dec;183(24):6991-8
references.pmid: 11717255
references.citation: Mol Microbiol. 2010 Jan;75(2):376-93
references.pmid: 19919668
references.citation: Mol Microbiol. 2005 Sep;57(6):1719-33
references.pmid: 16135236
references.citation: Mol Microbiol. 2002 Aug;45(4):967-82
references.pmid: 12180917
references.citation: Annu Rev Microbiol. 2000;54:735-74
references.pmid: 11018143
references.citation: Proc Natl Acad Sci U S A. 2004 Jan 6;101(1):70-5
references.pmid: 14694203
references.citation: Nature. 2008 Jan 24;451(7177):489-92
references.pmid: 18216859
references.citation: J Bacteriol. 2001 Feb;183(4):1159-67
references.pmid: 11157927
references.citation: Science. 1999 May 21;284(5418):1322-8
references.pmid: 10334981
references.citation: J Bacteriol. 2004 Jul;186(14):4645-54
references.pmid: 15231798
references.citation: Proc Natl Acad Sci U S A. 2007 Jan 9;104(2):485-90
references.pmid: 17202259
references.citation: Biophys J. 2003 Jan;84(1):571-7
references.pmid: 12524310
references.citation: Mol Microbiol. 1997 Nov;26(4):789-97
references.pmid: 9427408
references.citation: J Bacteriol. 2006 Oct;188(20):7039-48
references.pmid: 17015643
references.citation: Micron. 2007;38(3):291-301
references.pmid: 16920362
references.citation: Annu Rev Phytopathol. 2004;42:385-414
references.pmid: 15283671
references.citation: Ann Med. 2005;37(4):234-49
references.pmid: 16019722
references.citation: Annu Rev Microbiol. 2003;57:77-100
references.pmid: 12730325
references.citation: J Biol Chem. 2006 Jan 6;281(1):599-607
references.pmid: 16246841
references.citation: Nat Struct Mol Biol. 2009 May;16(5):477-85
references.pmid: 19396171
references.citation: Mol Microbiol. 1997 Mar;23(5):861-7
references.pmid: 9076724
references.citation: J Bacteriol. 1997 Feb;179(4):1307-16
references.pmid: 9023216
references.citation: J Bacteriol. 2009 May;191(9):3149-61
references.pmid: 19251862
references.citation: Mol Microbiol. 2001 Feb;39(3):652-63
references.pmid: 11169106
references.citation: Mol Microbiol. 2005 Mar;55(6):1658-70
references.pmid: 15752191
references.citation: J Bacteriol. 2009 Oct;191(19):6186-91
references.pmid: 19648242
references.citation: Nat Struct Mol Biol. 2007 Feb;14(2):131-7
references.pmid: 17237797
references.citation: J Bacteriol. 1997 Feb;179(3):813-7
references.pmid: 9006037
references.citation: Curr Opin Struct Biol. 2008 Apr;18(2):258-66
references.pmid: 18258424
references.citation: J Bacteriol. 2009 Jan;191(1):161-8
references.pmid: 18978053
references.citation: Infect Immun. 2004 Jul;72(7):4004-9
references.pmid: 15213145
references.citation: EMBO J. 2007 Jun 20;26(12):3015-24
references.pmid: 17510628
references.citation: J Cell Biol. 1999 Nov 1;147(3):683-93
references.pmid: 10545510
references.citation: Cell Microbiol. 2001 Dec;3(12):865-71
references.pmid: 11736997
references.citation: FEMS Microbiol Lett. 2000 May 1;186(1):85-90
references.pmid: 10779717
references.citation: Nature. 1994 Aug 25;370(6491):621-8
references.pmid: 8065448
references.citation: Proc Natl Acad Sci U S A. 2006 Aug 15;103(33):12529-33
references.pmid: 16888041
references.citation: Science. 2001 Dec 21;294(5551):2556-8
references.pmid: 11752577
references.citation: Microb Pathog. 1987 May;2(5):367-79
references.pmid: 3507556
references.citation: EMBO J. 2000 Aug 1;19(15):3876-87
references.pmid: 10921870
references.citation: Proc Natl Acad Sci U S A. 2000 Aug 29;97(18):10225-30
references.pmid: 10944190
references.citation: PLoS Pathog. 2010 Apr 01;6(4):e1000824
references.pmid: 20368966
references.citation: Nucleic Acids Res. 2007;35(20):e137
references.pmid: 17959646
references.citation: Nature. 2005 Oct 6;437(7060):911-5
references.pmid: 16208377
references.citation: Mol Microbiol. 1989 Oct;3(10):1455-9
references.pmid: 2693899
references.citation: Annu Rev Microbiol. 2006;60:425-49
references.pmid: 16753033
references.citation: Proc Natl Acad Sci U S A. 1992 Jul 1;89(13):5956-60
references.pmid: 1631080
references.citation: Proc Natl Acad Sci U S A. 2000 Sep 26;97(20):11008-13
references.pmid: 10984518
references.citation: EMBO J. 2006 Nov 15;25(22):5241-9
references.pmid: 17082772
references.citation: Nat Struct Mol Biol. 2009 May;16(5):468-76
references.pmid: 19396170
references.citation: Proc Natl Acad Sci U S A. 2001 Sep 25;98(20):11638-43
references.pmid: 11562461
references.citation: Mol Microbiol. 1995 Mar;15(6):1095-114
references.pmid: 7623665
references.citation: J Biol Chem. 2003 Jul 11;278(28):25816-24
references.pmid: 12734178
references.citation: Protein Sci. 2006 Apr;15(4):935-41
references.pmid: 16522800
references.citation: Nature. 2008 Jan 24;451(7177):485-8
references.pmid: 18216858
references.citation: Science. 1998 Apr 24;280(5363):602-5
references.pmid: 9554854
references.citation: J Bacteriol. 2006 May;188(10):3525-34
references.pmid: 16672607
references.citation: Nature. 2005 Jun 2;435(7042):702-7
references.pmid: 15931226
references.citation: J Mol Biol. 2000 May 12;298(4):577-83
references.pmid: 10788321
references.citation: J Biol Chem. 2003 May 9;278(19):17103-7
references.pmid: 12571230
references.citation: J Mol Biol. 1992 Jul 20;226(2):433-46
references.pmid: 1640458
references.citation: J Mol Biol. 1990 Feb 20;211(4):669-72
references.pmid: 2179562
references.citation: Biochemistry. 2006 Oct 3;45(39):11790-8
references.pmid: 17002279
references.citation: J Bacteriol. 2004 Aug;186(16):5366-75
references.pmid: 15292137
references.citation: Microbiology (Reading). 2008 Feb;154(Pt 2):593-607
references.pmid: 18227263
references.citation: J Bacteriol. 1994 Mar;176(6):1561-9
references.pmid: 8132449
references.citation: Gene. 1991 Dec 20;109(1):137-41
references.pmid: 1756974
references.citation: Science. 2005 Oct 28;310(5748):674-6
references.pmid: 16254184
references.citation: Mol Microbiol. 2006 Jul;61(1):119-25
references.pmid: 16824099