The RSC Chromatin Remodelling ATPase Translocates DNA with High Force and Small Step Size

Overview

Journal EMBO J

Specialties Biotechnology
Molecular Biology

Date 2011 May 10

PMID 21552204

Citations 58

Authors

George Sirinakis

Cedric R Clapier

Ying Gao

Ramya Viswanathan

Bradley R Cairns

Yongli Zhang

Affiliations

Soon will be listed here.

Abstract

ATP-dependent chromatin remodelling complexes use the energy of ATP hydrolysis to reposition and reconfigure nucleosomes. Despite their diverse functions, all remodellers share highly conserved ATPase domains, many shown to translocate DNA. Understanding remodelling requires biophysical knowledge of the DNA translocation process: how the ATPase moves DNA and generates force, and how translocation and force generation are coupled on nucleosomes. Here, we characterize the real-time activity of a minimal RSC translocase 'motor' on bare DNA, using high-resolution optical tweezers and a 'tethered' translocase system. We observe on dsDNA a processivity of ∼35 bp, a speed of ∼25 bp/s, and a step size of 2.0 (±0.4, s.e.m.) bp. Surprisingly, the motor is capable of moving against high force, up to 30 pN, making it one of the most force-resistant motors known. We also provide evidence for DNA 'buckling' at initiation. These observations reveal the ATPase as a powerful DNA translocating motor capable of disrupting DNA-histone interactions by mechanical force.

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