Dataset Information

Indentification of RecBCD Auxiliary ATP binding

ABSTRACT: The RecBCD helicase initiates double-stranded break repair in bacteria by processively unwinding DNA with a rate approaching ~1,600 bp·second, but the mechanism enabling such a fast rate is unknown. By using crosslinking followed by mass spectrometry, we revealed evidence for the existence of auxiliary binding sites in the RecC subunit. This was validated by equilibrium and time-resolved binding experiments, ensemble, and single-molecule unwinding assays. The essentiality and functionality of these sites are further demonstrated by their impact on the survival of E.coli after exposure to damage-inducing radiation.

INSTRUMENT(S): Q Exactive Plus

ORGANISM(S): Escherichia Coli

SUBMITTER: Oded Kleifeld

LAB HEAD: Arnon Henn

PROVIDER: PXD031540 | Pride | 2022-05-20

REPOSITORIES: Pride

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Auxiliary ATP binding sites support DNA unwinding by RecBCD.

Zananiri Rani R Mangapuram Venkata Sivasubramanyan S Gaydar Vera V Yahalom Dan D Malik Omri O Rudnizky Sergei S Kleifeld Oded O Kaplan Ariel A Henn Arnon A

Nature communications 20220404 1

The RecBCD helicase initiates double-stranded break repair in bacteria by processively unwinding DNA with a rate approaching ∼1,600 bp·s<sup>-1</sup>, but the mechanism enabling such a fast rate is unknown. Employing a wide range of methodologies - including equilibrium and time-resolved binding experiments, ensemble and single-molecule unwinding assays, and crosslinking followed by mass spectrometry - we reveal the existence of auxiliary binding sites in the RecC subunit, where ATP binds with l ...[more]

PMID: 35379800

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