PH modulates the binding of early growth response protein 1 transcription factor to DNA

David C. Mikles, Vikas Bhat, Brett J. Schuchardt, Brian J. Deegan, Kenneth L. Seldeen, Caleb B. McDonald, Amjad Farooq

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

The transcription factor early growth response protein (EGR)1 orchestrates a plethora of signaling cascades involved in cellular homeostasis, and its downregulation has been implicated in the development of prostate cancer. Herein, using a battery of biophysical tools, we show that the binding of EGR1 to DNA is tightly regulated by solution pH. Importantly, the binding affinity undergoes an enhancement of more than an order of magnitude with an increase in pH from 5 to 8, implying that the deprotonation of an ionizable residue accounts for such behavior. This ionizable residue is identified as His382 by virtue of the fact that its replacement by nonionizable residues abolishes the pH dependence of the binding of EGR1 to DNA. Notably, His382 inserts into the major groove of DNA, and stabilizes the EGR1-DNA interaction via both hydrogen bonding and van der Waals contacts. Remarkably, His382 is mainly conserved across other members of the EGR family, implying that histidine protonation-deprotonation may serve as a molecular switch for modulating the protein-DNA interactions that are central to this family of transcription factors. Collectively, our findings reveal an unexpected but a key step in the molecular recognition of the EGR family of transcription factors, and suggest that they may act as sensors of pH within the intracellular environment. We show that the binding of EGR1 transcription factor to DNA is tightly regulated by solution pH by virtue of the ability of His382 to undergo protonation- deprotonation equilibrium. Remarkably, His382 is predominantly conserved across other members of EGR1 family, implying that histidine protonation-deprotonation may serve as a molecular switch for modulating protein-DNA interactions central to this family of transcription factors.

Original languageEnglish
Pages (from-to)3669-3684
Number of pages16
JournalFEBS Journal
Volume280
Issue number15
DOIs
StatePublished - Aug 1 2013

Fingerprint

Early Growth Response Protein 1
Transcription Factors
Deprotonation
DNA
Protonation
Histidine
Proteins
Switches
Molecular recognition
Aptitude
Transcription factors
Hydrogen Bonding
Growth
Prostatic Neoplasms
Hydrogen bonds
Homeostasis
Down-Regulation

Keywords

  • histidine protonation
  • intracellular pH
  • protein dynamics
  • protein-DNA thermodynamics
  • zinc fingers

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Mikles, D. C., Bhat, V., Schuchardt, B. J., Deegan, B. J., Seldeen, K. L., McDonald, C. B., & Farooq, A. (2013). PH modulates the binding of early growth response protein 1 transcription factor to DNA. FEBS Journal, 280(15), 3669-3684. https://doi.org/10.1111/febs.12360

PH modulates the binding of early growth response protein 1 transcription factor to DNA. / Mikles, David C.; Bhat, Vikas; Schuchardt, Brett J.; Deegan, Brian J.; Seldeen, Kenneth L.; McDonald, Caleb B.; Farooq, Amjad.

In: FEBS Journal, Vol. 280, No. 15, 01.08.2013, p. 3669-3684.

Research output: Contribution to journalArticle

Mikles, DC, Bhat, V, Schuchardt, BJ, Deegan, BJ, Seldeen, KL, McDonald, CB & Farooq, A 2013, 'PH modulates the binding of early growth response protein 1 transcription factor to DNA', FEBS Journal, vol. 280, no. 15, pp. 3669-3684. https://doi.org/10.1111/febs.12360
Mikles DC, Bhat V, Schuchardt BJ, Deegan BJ, Seldeen KL, McDonald CB et al. PH modulates the binding of early growth response protein 1 transcription factor to DNA. FEBS Journal. 2013 Aug 1;280(15):3669-3684. https://doi.org/10.1111/febs.12360
Mikles, David C. ; Bhat, Vikas ; Schuchardt, Brett J. ; Deegan, Brian J. ; Seldeen, Kenneth L. ; McDonald, Caleb B. ; Farooq, Amjad. / PH modulates the binding of early growth response protein 1 transcription factor to DNA. In: FEBS Journal. 2013 ; Vol. 280, No. 15. pp. 3669-3684.
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