Kinetics and Thermodynamics of Phenotype: Unwinding and Rewinding the Nucleosome

JOURNAL OF MOLECULAR BIOLOGY

卷:423

Article

Mack, Andrew H.^1,2  Schlingman, Daniel J.^1,3  Ilagan, Robielyn P.³  Regan, Lynne^1,3,4  Mochrie, Simon G. J.^1,2,5 

[1] Yale Univ, Integrated Grad Program Phys & Engn Biol, New Haven, CT 06520 USA

[2] Yale Univ, Dept Appl Phys, New Haven, CT 06520 USA

[3] Yale Univ, Dept Mol Biophys & Biochem, New Haven, CT 06520 USA

[4] Yale Univ, Dept Chem, New Haven, CT 06520 USA

[5] Yale Univ, Dept Phys, New Haven, CT 06520 USA

关键词: chromatin;    single molecule;    optical tweezers;    free energy;    nucleosome;   

DOI  :  10.1016/j.jmb.2012.08.021

来源: Elsevier

【 摘 要 】

Chromatin remodeling is widely accepted as the mechanism that permits access to DNA by the transcription machinery. To date, however, there has been no experimental measurement of the changes in the kinetics and thermodynamics of the DNA histone octamer association that are required to remodel chromatin so that transcription may occur. Here, we present the results of optical tweezer measurements that compare the kinetic and thermodynamic properties of nucleosomes composed of unmodified histones with those of nucleosomes that contain a mutant histone H4 (H4-R45H), which has been shown to allow SWI/SNF remodeling factor-independent transcription from the yeast HO promoter in vivo. Our measurements, carried out in a force-clamp mode, determine the force-dependent unwinding and rewinding rates of the nucleosome inner turn. At each force studied, nucleosomes containing H4-R45H unwind more rapidly and rewind more slowly than nucleosomes containing unmodified H4, indicating that the latter are the more stable. Extrapolation to forces at which the winding and unwinding rates are equal determines the absolute free energy of the nucleosome inner turn to be -32k(B)T for nucleosomes containing unmodified H4 and -27k(B)T for nucleosomes containing H4-R45H. Thus, the loosening or remodeling caused by this point mutation, which is demonstrated to be sufficient to allow transcriptional machinery access to the HO promoter (in the absence of other remodeling factors), is 5k(B)T. The correlation between the free energy of the nucleosome inner turn and the sin (SWI/SNF-independent) transcription suggests that, beyond partial unwinding, complete histone unwinding may play a role in transcriptional activation. (C) 2012 Elsevier Ltd. All rights reserved.

【 授权许可】

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