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

• Published in: Journal of molecular recognition Vol. 23; no. 3; pp. 301 - 315
• Main Authors: Couturier, Marie, Buccellato, Matt, Costanzo, Stéphanie, Bourhis, Jean-Marie, Shu, Yaoling, Nicaise, Magali, Desmadril, Michel, Flaudrops, Christophe, Longhi, Sonia, Oglesbee, Michael
• Format: Journal Article
• Language: English
• Published: Chichester, UK John Wiley & Sons, Ltd 01.05.2010
• Subjects: Amino Acid Sequence; Animals; Binding Sites; Cell Line; hdj1; Hsp40; HSP40 Heat-Shock Proteins - chemistry; HSP40 Heat-Shock Proteins - genetics; HSP40 Heat-Shock Proteins - metabolism; Hsp70; HSP70 Heat-Shock Proteins - chemistry; HSP70 Heat-Shock Proteins - genetics; HSP70 Heat-Shock Proteins - metabolism; intrinsically disordered protein domain; Measles virus; Mice; Models, Molecular; nucleoprotein; Nucleoproteins - chemistry; Nucleoproteins - genetics; Nucleoproteins - metabolism; Protein Binding; Protein Structure, Tertiary; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Viral Proteins - chemistry; Viral Proteins - genetics; Viral Proteins - metabolism
• ISSN: 0952-3499; 1099-1352; 1099-1352
• DOI: 10.1002/jmr.982

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.