Role of Nhp6 protein in nucleosome unfolding by factor FACT
Abstract
The accessibility of DNA is important for the regulation of gene expression and provided by regulatory factors, such as FACT protein complex. As it was shown before, in the presence of Nhp6 protein, yeast FACT (yFACT) induces ATP-independent reversible unfolding of nucleosome, but the detailed mechanism of this process is unknown. In this study, we investigated whether presence of single Nhp6 molecule is sufficient for the unwinding of nucleosomal DNA by the yFACT factor, or whether the reorganization of the nucleosome structure requires the combined action of yFACT and several Nhp6 molecules. Analysis of the protein complex in the absence of nucleosomes by native gel electrophoresis has shown that yFACT itself can bind at least three Nhp6 molecules. Using single particle microscopy based on the Foerster resonance energy transfer, it is shown that with an increase in the ratio yFACT:Nhp6 from 1:10 to 1:1 at a constant concentration of Nhp6, the ability of yFACT to unfold nucleosomes does not increase, but decreases. Therefore, the unfolding of the nucleosome requires the binding of more than one Nhp6 molecule in the complex nucleosome:yFACT:Nhp6. The obtained data clarify the existing model of the reorganization of nucleosome structure by yFACT.
About the Authors
A. L. Sivkina
Lomonosov Moscow State University
Russian Federation
Russian Federation
Department of Bioengineering, School of Biology
Leninskiye gory 1–12, Moscow, 119234
A. V. Feofanov
Lomonosov Moscow State University; Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences
Russian Federation
Russian Federation
Department of Bioengineering, School of Biology
Leninskiye gory 1–12, Moscow, 119234;
Miklukho-Maklaya ul. 16/10, Moscow, 117997
M. P. Kirpichnikov
Lomonosov Moscow State University; Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences
Russian Federation
Russian Federation
Department of Bioengineering, School of Biology
Leninskiye gory 1–12, Moscow, 119234;
Miklukho-Maklaya ul. 16/10, Moscow, 117997
M. S. Akhtar
CSIR-Central Drug Research Institute
India
India
Biochemistry and Structural Biology Division
Sector 10, Jankipuram Extension, Sitapur Road, Lucknow, PIN 226031
V. M. Studitsky
Lomonosov Moscow State University; Fox Chase Cancer Center
Russian Federation
Russian Federation
Department of Bioengineering, School of Biology;
Leninskiye gory 1–12, Moscow, 119234, Russia;
333 Cottman Avenue, Philadelphia, PA 19111, USA
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Review
For citations:
Sivkina A.L., Feofanov A.V., Kirpichnikov M.P., Akhtar M.S., Studitsky V.M. Role of Nhp6 protein in nucleosome unfolding by factor FACT. Vestnik Moskovskogo universiteta. Seriya 16. Biologiya. 2021;76(4):213-218. (In Russ.)
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