title: The Hsp40 J-domain stimulates Hsp70 when tethered by the client to the ATPase domain.
abstract: The Escherichia coli Hsp40 DnaJ uses its J-domain (Jd) to couple ATP hydrolysis and client protein capture in Hsp70 DnaK. Fusion of the Jd to peptide p5 (as in Jdp5) dramatically increases the apparent affinity of the p5 moiety for DnaK in the presence of ATP, and Jdp5 stimulates ATP hydrolysis in DnaK by several orders of magnitude. NMR experiments with [(15)N]Jdp5 demonstrated that the peptide tethers the Jd to the ATPase domain. Thus, ATP hydrolysis and client protein binding in DnaK are coupled principally through the association of the client with DnaJ. Overexpression of a recombinant Jd was specifically toxic to cells that simultaneously expressed DnaK. No toxicity was observed when overexpressing Jdp5 or mutant Jd or when co-overexpressing the Jd and the nucleotide exchange factor GrpE. The results suggest that the Jd shifts DnaK to a client-bound form by stimulating the DnaK ATPase but only when the Jd is brought to DnaK by a client-Hsp40 complex.
authors.affiliation: Department of Biochemistry, Tulane University Health Sciences Center, New Orleans, Louisiana 70112, USA.
authors.forename: B Erin
authors.initials: BE
authors.lastname: Horne
authors.forename: Tingfeng
authors.initials: T
authors.lastname: Li
authors.forename: Pierre
authors.initials: P
authors.lastname: Genevaux
authors.forename: Costa
authors.initials: C
authors.lastname: Georgopoulos
authors.forename: Samuel J
authors.initials: SJ
authors.lastname: Landry
mesh_terms: D000251:Adenosine Triphosphatases / Q000737:chemistry*; D018833:Chaperonins / Q000737:chemistry; D004926:Escherichia coli / Q000201:enzymology*; D029968:Escherichia coli Proteins / Q000737:chemistry; D050956:HSP40 Heat-Shock Proteins / Q000737:chemistry*; D018840:HSP70 Heat-Shock Proteins / Q000737:chemistry*; D006868:Hydrolysis; D009682:Magnetic Resonance Spectroscopy; D018832:Molecular Chaperones; D009154:Mutation; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry
chemical_list: D029968:Escherichia coli Proteins; D050956:HSP40 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins; D018832:Molecular Chaperones; D011994:Recombinant Proteins; D000251:Adenosine Triphosphatases; D018833:Chaperonins; C063842:dnaK protein, E coli
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