Interaction of MxiG with the cytosolic complex of the type III secretion system controls Shigella virulence

被引:15
作者
Barison, Nicola [1 ]
Lambers, Jutta [1 ]
Hurwitz, Robert [2 ]
Kolbe, Michael [1 ]
机构
[1] Max Planck Inst Infect Biol, Dept Cellular Microbiol, D-10117 Berlin, Germany
[2] Max Planck Inst Infect Biol, Prot Purificat Core Facil, D-10117 Berlin, Germany
关键词
FHA domain; protein phosphorylation; Spa33; protein transport; EPITHELIAL-CELLS; NEEDLE COMPLEX; PROTEIN SECRETION; FLEXNERI; IPAB; MODEL; CONSERVATION; INVASINS; RELEASE; SIGNAL;
D O I
10.1096/fj.11-197160
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Gram-negative bacteria use the type 3 secretion system (T3SS) to colonize host cells. T3SSs are ring-shaped macromolecular complexes specific for the transport of effector molecules into host cells. It was recently suggested that a cytosolic ring-shaped protein complex delivers effector molecules to the T3SS. However, how transport of effector proteins is regulated is not known. Here, we report the high-resolution X-ray crystal structure of the whole cytosolic domain of MxiG (MxiG(1-126)), a major component of the inner T3SS rings in Shigella flexneri. MxiG(1-126) folds as an FHA domain, which specifically binds phosphorylated threonines. Indeed, MxiG(1-126) binds to Spa33, a cytoplasmic-ring component of Shigella, as revealed in pulldown studies. Surface plasmon resonance analysis showed specific interaction of MxiG with a Spa33 peptide only if phosphorylated. In total, 24 copies of the MxiG(1-126) crystal structure were fitted into the cryo-EM map of the Shigella T3SS. The phosphoprotein binding site of each MxiG molecule faces the channel of the T3SS, allowing interaction with cytosolic binding partners. Secretion assays and host cell invasion studies of complemented Shigella knockout cells indicated that the phosphoprotein binding of MxiG is essential for bacterial virulence. Our findings suggest that MxiG is involved in T3SS regulation.-Barison, N., Lambers, J., Hurwitz, R., Kolbe, M. Interaction of MxiG with the cytosolic complex of the type III secretion system controls Shigella virulence. FASEB J. 26, 1717-1726 (2012). www.fasebj.org
引用
收藏
页码:1717 / 1726
页数:10
相关论文
共 51 条
[1]   Chaperone release and unfolding of substrates in type III secretion [J].
Akeda, Y ;
Galán, JE .
NATURE, 2005, 437 (7060) :911-915
[2]   Genetic analysis of the Salmonella enterica type III secretion-associated ATPase InvC defines discrete functional domains [J].
Akeda, Y ;
Galán, JE .
JOURNAL OF BACTERIOLOGY, 2004, 186 (08) :2402-2412
[3]   Identification of Vibrio cholerae Type III Secretion System Effector Proteins [J].
Alam, Ashfaqul ;
Miller, Kelly A. ;
Chaand, Mudit ;
Butler, J. Scott ;
Dziejman, Michelle .
INFECTION AND IMMUNITY, 2011, 79 (04) :1728-1740
[4]   Molecular structure of EmbR, a response element of Ser/Thr kinase signaling in Mycobacterium tuberculosis [J].
Alderwick, LJ ;
Molle, V ;
Kremer, L ;
Cozzone, AJ ;
Dafforn, TR ;
Besra, GS ;
Fütterer, K .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2006, 103 (08) :2558-2563
[5]   MXIG, A MEMBRANE-PROTEIN REQUIRED FOR SECRETION OF SHIGELLA SPP LPA INVASINS - INVOLVEMENT IN ENTRY INTO EPITHELIAL-CELLS AND IN INTERCELLULAR DISSEMINATION [J].
ALLAOUI, A ;
SANSONETTI, PJ ;
MENARD, R ;
BARZU, S ;
MOUNIER, J ;
PHALIPON, A ;
PARSOT, C .
MOLECULAR MICROBIOLOGY, 1995, 17 (03) :461-470
[6]   Characterization of the Yersinia enterocolitica type III secretion ATPase YscN and its regulator, YscL [J].
Blaylock, Bill ;
Riordan, Kelly E. ;
Missiakas, Dominique M. ;
Schneewind, Olaf .
JOURNAL OF BACTERIOLOGY, 2006, 188 (10) :3525-3534
[7]   The tripartite type III secreton of Shigella flexneri inserts IpaB and IpaC into host membranes [J].
Blocker, A ;
Gounon, P ;
Larquet, E ;
Niebuhr, K ;
Cabiaux, V ;
Parsot, C ;
Sansonetti, P .
JOURNAL OF CELL BIOLOGY, 1999, 147 (03) :683-693
[8]   Structure and composition of the Shigella flexneri 'needle complex', a part of its type III secreton [J].
Blocker, A ;
Jouihri, N ;
Larquet, E ;
Gounon, P ;
Ebel, F ;
Parsot, C ;
Sansonetti, P ;
Allaoui, A .
MOLECULAR MICROBIOLOGY, 2001, 39 (03) :652-663
[9]   What's the point of the type III secretion system needle? [J].
Blocker, Ariel J. ;
Deane, Janet E. ;
Veenendaal, Andreas K. J. ;
Roversi, Pietro ;
Hodgkinson, Julie L. ;
Johnson, Steven ;
Lea, Susan M. .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2008, 105 (18) :6507-6513
[10]   The 33 carboxyl-terminal residues of Spa40 orchestrate the multi-step assembly process of the type III secretion needle complex in Shigella flexneri [J].
Botteaux, Anne ;
Kayath, Christian A. ;
Page, Anne-Laure ;
Jouihri, Nouredine ;
Sani, Musa ;
Boekema, Egbert ;
Biskri, Latefa ;
Parsot, Claude ;
Allaoui, Abdelmounaaim .
MICROBIOLOGY-SGM, 2010, 156 :2807-2817