Large-scale ATP-independent nucleosome unfolding by a histone chaperone.
DNA accessibility to regulatory proteins is substantially influenced by nucleosome structure and dynamics. The facilitates chromatin transcription (FACT) complex increases the accessibility of nucleosomal DNA, but the mechanism and extent of its nucleosome reorganization activity are unknown. Here we determined the effects of FACT from the yeast Saccharomyces cerevisiae on single nucleosomes by using single-particle Förster resonance energy transfer (spFRET) microscopy. FACT binding results in dramatic ATP-independent, symmetrical and reversible DNA uncoiling that affects at least 70% of the DNA within a nucleosome, occurs without apparent loss of histones and proceeds via an 'all-or-none' mechanism. A mutated version of FACT is defective in uncoiling, and a histone mutation that suppresses phenotypes caused by this FACT mutation in vivo restores the uncoiling activity in vitro. Thus, FACT-dependent nucleosome unfolding modulates the accessibility of nucleosomal DNA, and this activity is an important function of FACT in vivo.
Biology Faculty, Lomonosov, Moscow State University, Moscow, Russia.
Maria E
ME
Valieva
Biology Faculty, Lomonosov, Moscow State University, Moscow, Russia.
Grigoriy A
GA
Armeev
Biology Faculty, Lomonosov, Moscow State University, Moscow, Russia.|Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russia.
Kseniya S
KS
Kudryashova
Biology Faculty, Lomonosov, Moscow State University, Moscow, Russia.
Nadezhda S
NS
Gerasimova
Biology Faculty, Lomonosov, Moscow State University, Moscow, Russia.
Alexey K
AK
Shaytan
Biology Faculty, Lomonosov, Moscow State University, Moscow, Russia.|Fox Chase Cancer Center, Philadelphia, Pennsylvania, USA.
Olga I
OI
Kulaeva
Department of Biochemistry, University of Utah School of Medicine, Salt Lake City, Utah, USA.
Laura L
LL
McCullough
Department of Biochemistry, University of Utah School of Medicine, Salt Lake City, Utah, USA.
Tim
T
Formosa
Institute of Gene Biology, Russian Academy of Sciences, Moscow, Russia.
Pavel G
PG
Georgiev
Biology Faculty, Lomonosov, Moscow State University, Moscow, Russia.|Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russia.
Mikhail P
MP
Kirpichnikov
Biology Faculty, Lomonosov, Moscow State University, Moscow, Russia.|Fox Chase Cancer Center, Philadelphia, Pennsylvania, USA.|Institute of Gene Biology, Russian Academy of Sciences, Moscow, Russia.
Vasily M
VM
Studitsky
Biology Faculty, Lomonosov, Moscow State University, Moscow, Russia.|Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russia.
Alexey V
AV
Feofanov
D000255:Adenosine Triphosphate / Q000378:metabolism*; D004271:DNA, Fungal / Q000737:chemistry / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D031541:Fluorescence Resonance Energy Transfer; D006609:High Mobility Group Proteins / Q000737:chemistry / Q000378:metabolism*; D006657:Histones / Q000737:chemistry / Q000378:metabolism*; D008958:Models, Molecular; D018832:Molecular Chaperones / Q000737:chemistry / Q000378:metabolism; D009690:Nucleic Acid Conformation; D009707:Nucleosomes / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000378:metabolism*; D014157:Transcription Factors / Q000737:chemistry / Q000378:metabolism*; D035602:Transcriptional Elongation Factors / Q000737:chemistry / Q000378:metabolism*
D004271:DNA, Fungal; D004268:DNA-Binding Proteins; C541535:FACT protein, S cerevisiae; D006609:High Mobility Group Proteins; D006657:Histones; D018832:Molecular Chaperones; D009707:Nucleosomes; C106768:POB3 protein, S cerevisiae; C071238:SPT16 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D035602:Transcriptional Elongation Factors; D000255:Adenosine Triphosphate