High affinity binding between Hsp70 and the C-terminal domain of the measles virus nucleoprotein requires an Hsp40 co-chaperone

Authors

  • Marie Couturier,

    1. Architecture et Fonction des Macromolécules Biologiques, UMR 6098 CNRS et Universités Aix-Marseille I et II, Campus de Luminy, 13288 Marseille, Cedex 9, France
    Search for more papers by this author
  • Matt Buccellato,

    1. Department of Veterinary Biosciences, The Ohio State University, 1925 Coffey Road, Columbus, OH 43210, USA
    Search for more papers by this author
  • Stéphanie Costanzo,

    1. Architecture et Fonction des Macromolécules Biologiques, UMR 6098 CNRS et Universités Aix-Marseille I et II, Campus de Luminy, 13288 Marseille, Cedex 9, France
    Search for more papers by this author
  • Jean-Marie Bourhis,

    1. Architecture et Fonction des Macromolécules Biologiques, UMR 6098 CNRS et Universités Aix-Marseille I et II, Campus de Luminy, 13288 Marseille, Cedex 9, France
    2. IBCP UMR 5086 CNRS, 7, passage du Vercors 69367 Lyon, Cedex 07, France
    Search for more papers by this author
  • Yaoling Shu,

    1. Department of Veterinary Biosciences, The Ohio State University, 1925 Coffey Road, Columbus, OH 43210, USA
    Search for more papers by this author
  • Magali Nicaise,

    1. Institut de Biochimie et Biophysique Moléculaire et Cellulaire, UMR 8619 CNRS, Université de Paris-Sud, Bât 430, 91405 Orsay, Cedex, France
    Search for more papers by this author
  • Michel Desmadril,

    1. Institut de Biochimie et Biophysique Moléculaire et Cellulaire, UMR 8619 CNRS, Université de Paris-Sud, Bât 430, 91405 Orsay, Cedex, France
    Search for more papers by this author
  • Christophe Flaudrops,

    1. Architecture et Fonction des Macromolécules Biologiques, UMR 6098 CNRS et Universités Aix-Marseille I et II, Campus de Luminy, 13288 Marseille, Cedex 9, France
    2. Laboratoire de Bactériologie et de Virologie, CHU La Timone, 365 Rue Saint Pierre, 13385 Marseille, Cedex 5, France
    Search for more papers by this author
  • Sonia Longhi,

    Corresponding author
    1. Architecture et Fonction des Macromolécules Biologiques, UMR 6098 CNRS et Universités Aix-Marseille I et II, Campus de Luminy, 13288 Marseille, Cedex 9, France
    • AFMB, UMR 6098, 163, Avenue de Luminy, Case 932, 13288 Marseille, Cedex 09, France.
    Search for more papers by this author
  • Michael Oglesbee

    Corresponding author
    1. Department of Veterinary Biosciences, The Ohio State University, 1925 Coffey Road, Columbus, OH 43210, USA
    • Department Veterinary Biosciences, The Ohio State University, 1925 Coffey Road, Columbus, OH 43210, USA.
    Search for more papers by this author

Abstract

The major inducible 70 kDa heat shock protein (hsp70) binds the measles virus (MeV) nucleocapsid with high affinity in an ATP-dependent manner, stimulating viral transcription and genome replication, and profoundly influencing virulence in mouse models of brain infection. Binding is mediated by two hydrophobic motifs (Box-2 and Box-3) located within the C-terminal domain (NTAIL) of the nucleocapsid protein, with NTAIL being an intrinsically disordered domain. The current work showed that high affinity hsp70 binding to NTAIL requires an hsp40 co-chaperone that interacts primarily with the hsp70 nucleotide binding domain (NBD) and displays no significant affinity for NTAIL. Hsp40 directly enhanced hsp70 ATPase activity in an NTAIL-dependent manner, and formation of hsp40–hsp70–NTAIL intracellular complexes required the presence of NTAIL Box-2 and 3. Results are consistent with the functional interplay between hsp70 nucleotide and substrate binding domains (SBD), where ATP hydrolysis is rate limiting to high affinity binding to client proteins and is enhanced by hsp40. As such, hsp40 is an essential variable in understanding the outcome of MeV–hsp70 interactions. Copyright © 2009 John Wiley & Sons, Ltd.

Ancillary