Allosteric regulation of the primase (DnaG) activity by the clamp-loader (τ) in vitro

Kiran Chintakayala, Cristina MacHón, Anna Haroniti, Marilyn A. Larson, Steven H. Hinrichs, Mark A. Griep, Panos Soultanas

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

During DNA replication the helicase (DnaB) recruits the primase (DnaG) in the replisome to initiate the polymerization of new DNA strands. DnaB is attached to the τ subunit of the clamp-loader that loads the β clamp and interconnects the core polymerases on the leading and lagging strands. The τ-DnaB-DnaG ternary complex is at the heart of the replisome and its function is likely to be modulated by a complex network of allosteric interactions. Using a stable ternary complex comprising the primase and helicase from Geobacillus stearothermophilus and the τ subunit of the clamp-loader from Bacillus subtilis we show that changes in the DnaB-τ interaction can stimulate allosterically primer synthesis by DnaG in vitro. The A550V τ mutant stimulates the primase activity more efficiently than the native protein. Truncation of the last 18 C-terminal residues of τ elicits a DnaG-stimulatory effect in vitro that appears to be suppressed in the native τ protein. Thus changes in the τ-DnaB interaction allosterically affect primer synthesis. Although these C-terminal residues of τ are not involved directly in the interaction with DnaB, they may act as a functional gateway for regulation of primer synthesis by τ-interacting components of the replisome through the τ-DnaB-DnaG pathway.

Original languageEnglish (US)
Pages (from-to)537-549
Number of pages13
JournalMolecular Microbiology
Volume72
Issue number2
DOIs
StatePublished - Apr 1 2009

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Allosteric Regulation
DNA Primase
DNA Helicases
Geobacillus stearothermophilus
Bacillus subtilis
DNA Replication
Polymerization
Proteins
DNA
In Vitro Techniques

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

Cite this

Allosteric regulation of the primase (DnaG) activity by the clamp-loader (τ) in vitro. / Chintakayala, Kiran; MacHón, Cristina; Haroniti, Anna; Larson, Marilyn A.; Hinrichs, Steven H.; Griep, Mark A.; Soultanas, Panos.

In: Molecular Microbiology, Vol. 72, No. 2, 01.04.2009, p. 537-549.

Research output: Contribution to journalArticle

Chintakayala, Kiran ; MacHón, Cristina ; Haroniti, Anna ; Larson, Marilyn A. ; Hinrichs, Steven H. ; Griep, Mark A. ; Soultanas, Panos. / Allosteric regulation of the primase (DnaG) activity by the clamp-loader (τ) in vitro. In: Molecular Microbiology. 2009 ; Vol. 72, No. 2. pp. 537-549.
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