Legionella effector MavC targets the Ube2N∼Ub conjugate for noncanonical ubiquitination

被引:27
作者
Puvar, Kedar [1 ]
Iyer, Shalini [1 ]
Fu, Jiaqi [2 ,3 ]
Kenny, Sebastian [1 ]
Teron, Kristos I. Negron [1 ]
Luo, Zhao-Qing [2 ,3 ]
Brzovic, Peter S. [4 ]
Klevit, Rachel E. [4 ]
Das, Chittaranjan [1 ]
机构
[1] Purdue Univ, Dept Chem, W Lafayette, IN 47907 USA
[2] Purdue Univ, Purdue Inst Inflammat Immunol & Infect Dis, W Lafayette, IN 47907 USA
[3] Purdue Univ, Dept Biol Sci, W Lafayette, IN 47907 USA
[4] Univ Washington, Dept Biochem, Seattle, WA 98195 USA
关键词
STRUCTURAL BASIS; E2; ENZYMES; BACTERIAL; SYSTEM; MANIPULATION; DEAMIDATION; RECOGNITION; FAMILY; LIGASE; UBC13;
D O I
10.1038/s41467-020-16211-x
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
The bacterial effector MavC modulates the host immune response by blocking Ube2N activity employing an E1-independent ubiquitin ligation, catalyzing formation of a gamma-glutamyl-epsilon-Lys (Gln40(Ub)-Lys92(Ube2N)) isopeptide crosslink using a transglutaminase mechanism. Here we provide biochemical evidence in support of MavC targeting the activated, thioester-linked Ube2N similar to ubiquitin conjugate, catalyzing an intramolecular transglutamination reaction, covalently crosslinking the Ube2N and Ub subunits effectively inactivating the E2 similar to Ub conjugate. Ubiquitin exhibits weak binding to MavC alone, but shows an increase in affinity when tethered to Ube2N in a disulfide-linked substrate that mimics the charged E2 similar to Ub conjugate. Crystal structures of MavC in complex with the substrate mimic and crosslinked product provide insights into the reaction mechanism and underlying protein dynamics that favor transamidation over deamidation, while revealing a crucial role for the structurally unique insertion domain in substrate recognition. This work provides a structural basis of ubiquitination by transglutamination and identifies this enzyme's true physiological substrate. The Legionella pneumophila effector MavC inhibits the human ubiquitin-conjugating enzyme Ube2N. Here, the authors combine NMR, X-ray crystallography and biochemical assays and show that MavC catalyses the intramolecular transglutaminase reaction between the Ube2N and Ub subunits of the Ube2N similar to Ub conjugate and present the substrate- and product-bound MavC crystal structures.
引用
收藏
页数:14
相关论文
共 53 条
  • [1] PHENIX: a comprehensive Python']Python-based system for macromolecular structure solution
    Adams, Paul D.
    Afonine, Pavel V.
    Bunkoczi, Gabor
    Chen, Vincent B.
    Davis, Ian W.
    Echols, Nathaniel
    Headd, Jeffrey J.
    Hung, Li-Wei
    Kapral, Gary J.
    Grosse-Kunstleve, Ralf W.
    McCoy, Airlie J.
    Moriarty, Nigel W.
    Oeffner, Robert
    Read, Randy J.
    Richardson, David C.
    Richardson, Jane S.
    Terwilliger, Thomas C.
    Zwart, Peter H.
    [J]. ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY, 2010, 66 : 213 - 221
  • [2] Exploitation of eukaryotic ubiquitin signaling pathways by effectors translocated by bacterial type III and type IV secretion systems
    Angot, Aurelie
    Vergunst, Annette
    Genin, Stephane
    Peeters, Nemo
    [J]. PLOS PATHOGENS, 2007, 3 (01) : 1 - 13
  • [3] THE CCP4 SUITE - PROGRAMS FOR PROTEIN CRYSTALLOGRAPHY
    BAILEY, S
    [J]. ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY, 1994, 50 : 760 - 763
  • [4] 2 DISTINCT DEFECTS IN INTRACELLULAR GROWTH COMPLEMENTED BY A SINGLE GENETIC-LOCUS IN LEGIONELLA-PNEUMOPHILA
    BERGER, KH
    ISBERG, RR
    [J]. MOLECULAR MICROBIOLOGY, 1993, 7 (01) : 7 - 19
  • [5] Stereospecific interactions of proline residues in protein structures and complexes
    Bhattacharyya, R
    Chakrabarti, P
    [J]. JOURNAL OF MOLECULAR BIOLOGY, 2003, 331 (04) : 925 - 940
  • [6] Manipulation of host-cell pathways by bacterial pathogens
    Bhavsar, Amit P.
    Guttman, Julian A.
    Finlay, B. Brett
    [J]. NATURE, 2007, 449 (7164) : 827 - 834
  • [7] Phosphoribosylation of Ubiquitin Promotes Serine Ubiquitination and Impairs Conventional Ubiquitination
    Bhogaraju, Sagar
    Kalayil, Sissy
    Liu, Yaobin
    Bonn, Florian
    Colby, Thomas
    Matic, Ivan
    Dikic, Ivan
    [J]. CELL, 2016, 167 (06) : 1636 - +
  • [8] Genomic analysis of 38 Legionella species identifies large and diverse effector repertoires
    Burstein, David
    Amaro, Francisco
    Zusman, Tal
    Lifshitz, Ziv
    Cohen, Ofir
    Gilbert, Jack A.
    Pupko, Tal
    Shuman, Howard A.
    Segal, Gil
    [J]. NATURE GENETICS, 2016, 48 (02) : 167 - 175
  • [9] MolProbity: all-atom structure validation for macromolecular crystallography
    Chen, Vincent B.
    Arendall, W. Bryan, III
    Headd, Jeffrey J.
    Keedy, Daniel A.
    Immormino, Robert M.
    Kapral, Gary J.
    Murray, Laura W.
    Richardson, Jane S.
    Richardson, David C.
    [J]. ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY, 2010, 66 : 12 - 21
  • [10] The demographics of the ubiquitin system
    Clague, Michael J.
    Heride, Claire
    Urbe, Sylvie
    [J]. TRENDS IN CELL BIOLOGY, 2015, 25 (07) : 417 - 426