Thermodynamics and Kinetics of Hsp70 Association with A + U-rich mRNA-destabilizing Sequences

Gerald M. Wilson, Kristina Sutphen, Sandhya Bolikal, Keng Yu Chuang, Gary Brewer

Research output: Contribution to journalArticle

56 Citations (Scopus)

Abstract

Rapid mRNA degradation directed by A + U-rich elements (AREs) is mediated by the interaction of specific RNA-binding proteins to these sequences. The protein chaperone Hsp70 has been identified in a cellular complex containing the ARE-binding protein AUF1 and has also been detected in direct contact with A + U-rich RNA substrates, indicating that Hsp70 may be involved in the regulation of ARE-directed mRNA turnover. By using gel mobility shift and fluorescence anisotropy assays, we have determined that Hsp70 directly and specifically associates with U-rich RNA substrates in solution. With the ARE from tumor necrosis factor α (TNFα) mRNA, Hsp70 forms a dynamic complex consistent with a 1:1 association of protein:RNA but demonstrates cooperative binding behavior on polyuridylate substrates. Unlike AUF1, the RNA binding activity of Hsp70 is not regulated by ion-dependent folding of the TNFα ARE, suggesting that AUF1 and Hsp70 recognize distinct binding determinants on this RNA substrate. Binding of Hsp70 to the TNFα ARE is driven entirely by enthalpy at physiological temperatures, indicating that burial of hydrophobic surfaces is likely the principal mechanism stabilizing the Hsp70·RNA complex. Potential roles for the interaction of Hsp70 with ARE-containing mRNAs in the regulation of mRNA turnover and/or translational efficiency are discussed.

Original languageEnglish (US)
Pages (from-to)44450-44456
Number of pages7
JournalJournal of Biological Chemistry
Volume276
Issue number48
DOIs
StatePublished - Nov 30 2001

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Thermodynamics
RNA
Messenger RNA
Kinetics
Tumor Necrosis Factor-alpha
Substrates
Burial
Fluorescence Polarization
RNA-Binding Proteins
RNA Stability
Cooperative Behavior
Enthalpy
Assays
Carrier Proteins
Proteins
Anisotropy
Gels
Fluorescence
Association reactions
Ions

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Wilson, Gerald M. ; Sutphen, Kristina ; Bolikal, Sandhya ; Chuang, Keng Yu ; Brewer, Gary. / Thermodynamics and Kinetics of Hsp70 Association with A + U-rich mRNA-destabilizing Sequences. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 48. pp. 44450-44456.
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Thermodynamics and Kinetics of Hsp70 Association with A + U-rich mRNA-destabilizing Sequences. / Wilson, Gerald M.; Sutphen, Kristina; Bolikal, Sandhya; Chuang, Keng Yu; Brewer, Gary.

In: Journal of Biological Chemistry, Vol. 276, No. 48, 30.11.2001, p. 44450-44456.

Research output: Contribution to journalArticle

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