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The molecular basis of ubiquitin-like protein NEDD8 deamidation by the bacterial effector protein Cif

DOI: 10.1073/pnas.1112107109 DOI Help

Authors: A. Crow (John Innes Centre, Norwich Research Park) , R. K. Hughes (John Innes Centre, Norwich Research Park) , F. Taieb (INP-ENV de Toulouse, Institut National de la Recherche Agronomique Toulouse, Institut National de la Santé et de la Recherche Médicale Toulouse, Université Paul Sabatier de Toulousee) , E. Oswald (erche Agronomique Toulouse, Institut National de la Santé et de la Recherche Médicale Toulouse, Université Paul Sabatier de Toulouse, Centre National de la Recherche Scientifique, Centre Hospitalier Universitaire de Toulouse) , M. J. Banfield (John Innes Centre, Norwich Research Park)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Proceedings Of The National Academy Of Sciences , VOL 109 (27) , PAGES E1830 - E1838

State: Published (Approved)
Published: July 2012
Diamond Proposal Number(s): 7641

Abstract: The cycle inhibiting factors (Cifs) are a family of translocated effector proteins, found in diverse pathogenic bacteria, that interfere with the host cell cycle by catalyzing the deamidation of a specific glutamine residue (Gln40) in NEDD8 and the related protein ubiquitin. This modification prevents recycling of neddylated cullin-RING ligases, leading to stabilization of various cullin-RING ligase targets, and also prevents polyubiquitin chain formation. Here, we report the crystal structures of two Cif/NEDD8 complexes, revealing a conserved molecular interface that defines enzyme/substrate recognition. Mutation of residues forming the interface suggests that shape complementarity, rather than specific individual interactions, is a critical feature for complex formation. We show that Cifs from diverse bacteria bind NEDD8 in vitro and conclude that they will all interact with their substrates in the same way. The “occluding loop” in Cif gates access to Gln40 by forcing a conformational change in the C terminus of NEDD8. We used native PAGE to follow the activity of Cif from the human pathogen Yersinia pseudotuberculosis and selected variants, and the position of Gln40 in the active site has allowed us to propose a catalytic mechanism for these enzymes.

Journal Keywords: bacterial pathogenesis; cyclomodulins; host cell manipulation; structural biology; type III secreted effector proteins

Subject Areas: Biology and Bio-materials, Chemistry


Instruments: I02-Macromolecular Crystallography , I24-Microfocus Macromolecular Crystallography