A zinc finger-like domain of the molecular chaperone DnaJ is involved in binding to denatured protein substrates

Alexander Szabo, Richard Korszun, F. Ulrich Hartl, John Flanagan

Research output: Contribution to journalArticle

259 Citations (Scopus)

Abstract

The Escherichia coil heat-shock protein DnaJ cooperates with the Hsp70 homolog DnaK in protein folding in vitro and in vivo. Little is known about the structural features of DnaJ that mediate its interaction with DnaK and unfolded polypeptide. DnaJ contains at least four blocks of sequence representing potential functional domains which have been conserved throughout evolution. In order to understand the role of each of these regions, we have analyzed DnaJ fragments in reactions corresponding to known functions of the intact protein. Both the N-terminal 70 amino acid 'J-domain' and a 35 amino acid glycine-phenylalanine region following it are required for interactions with DnaK. However, only complete DnaJ can cooperate with DnaK and a third protein, GrpE, in refolding denatured firefly luciferase. As demonstrated by atomic absorption and extended X-ray absorption fine structure spectroscopy (EXAFS), the 90 amino acid cysteine-rich region of DnaJ contains two Zn atoms tetrahedrally coordinated to four cysteine residues, resembling their arrangement in the C4 Zn binding domains of certain DNA binding proteins. Interestingly, binding experiments and cross-linking studies indicate that this Zn finger-like domain is required for the DnaJ molecular chaperone to specifically recognize and bind to proteins in their denatured state.

Original languageEnglish (US)
Pages (from-to)408-417
Number of pages10
JournalEMBO Journal
Volume15
Issue number2
StatePublished - Jan 15 1996

Fingerprint

Molecular Chaperones
Zinc Fingers
Zinc
Amino Acids
Cysteine
Substrates
Extended X ray absorption fine structure spectroscopy
Firefly Luciferases
Protein folding
Escherichia
Proteins
Protein Folding
DNA-Binding Proteins
Heat-Shock Proteins
Phenylalanine
Glycine
Spectrum Analysis
X-Rays
Atoms
Peptides

All Science Journal Classification (ASJC) codes

  • Neuroscience(all)
  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Cite this

Szabo, Alexander ; Korszun, Richard ; Hartl, F. Ulrich ; Flanagan, John. / A zinc finger-like domain of the molecular chaperone DnaJ is involved in binding to denatured protein substrates. In: EMBO Journal. 1996 ; Vol. 15, No. 2. pp. 408-417.
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A zinc finger-like domain of the molecular chaperone DnaJ is involved in binding to denatured protein substrates. / Szabo, Alexander; Korszun, Richard; Hartl, F. Ulrich; Flanagan, John.

In: EMBO Journal, Vol. 15, No. 2, 15.01.1996, p. 408-417.

Research output: Contribution to journalArticle

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