Helix-destabilizing properties of the human single-stranded DNA- and RNA-binding protein purα

Nune Darbinian, Gary L. Gallia, Kamel Khalili

Research output: Contribution to journalArticlepeer-review

Abstract

Purα is a ubiquitous nucleic acid-binding protein which has been implicated in the control of eukaryotic gene transcription. Further, Purα associates with DNA sequences positioned in close proximity to viral and cellular origins of replication suggesting a role for this protein in DNA replication. As initiation of transcription and replication require alteration in the structure of duplex DNA, we investigated the DNA unwinding activity of this single-stranded nucleic acid-binding protein. Here we demonstrate that Puree has the ability to displace an oligonucleotide annealed to single-stranded M13 DNA. The helix unwinding activity of Puree was dose-, time- and temperature-dependent and ATP-independent. Results from mapping studies revealed that the central region of Purα, spanning amino acids 72-274, was important for the helix-destabilizing activity of this protein. The region of Puree which was involved in the helix-destabilizing activity mapped to the DNA-binding domain of this protein. Results from heat inactivation experiments demonstrated that the helix-destabilizing activity of Puree correlates with its capacity to interact with DNA containing the PUR element. Taken together, these studies demonstrate that Puree possesses helix-destabilizing activity and that this activity maps to and correlates with its ability to interact with DNA.

Original languageEnglish (US)
Pages (from-to)589-595
Number of pages7
JournalJournal of cellular biochemistry
Volume80
Issue number4
DOIs
StatePublished - Mar 15 2001

Keywords

  • Helix-destabilization
  • Nucleic acid
  • Purα
  • Replication
  • Single-stranded
  • Transc ription
  • Unwinding

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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