TY - JOUR
T1 - Electron microscopy analysis of ATP-independent nucleosome unfolding by FACT
AU - Sivkina, Anastasiia L.
AU - Karlova, Maria G.
AU - Valieva, Maria E.
AU - McCullough, Laura L.
AU - Formosa, Timothy
AU - Shaytan, Alexey K.
AU - Feofanov, Alexey V.
AU - Kirpichnikov, Mikhail P.
AU - Sokolova, Olga S.
AU - Studitsky, Vasily M.
N1 - Publisher Copyright:
© 2022, The Author(s).
PY - 2022/12
Y1 - 2022/12
N2 - FACT is a histone chaperone that participates in nucleosome removal and reassembly during transcription and replication. We used electron microscopy to study FACT, FACT:Nhp6 and FACT:Nhp6:nucleosome complexes, and found that all complexes adopt broad ranges of configurations, indicating high flexibility. We found unexpectedly that the DNA binding protein Nhp6 also binds to the C-terminal tails of FACT subunits, inducing more open geometries of FACT even in the absence of nucleosomes. Nhp6 therefore supports nucleosome unfolding by altering both the structure of FACT and the properties of nucleosomes. Complexes formed with FACT, Nhp6, and nucleosomes also produced a broad range of structures, revealing a large number of potential intermediates along a proposed unfolding pathway. The data suggest that Nhp6 has multiple roles before and during nucleosome unfolding by FACT, and that the process proceeds through a series of energetically similar intermediate structures, ultimately leading to an extensively unfolded form.
AB - FACT is a histone chaperone that participates in nucleosome removal and reassembly during transcription and replication. We used electron microscopy to study FACT, FACT:Nhp6 and FACT:Nhp6:nucleosome complexes, and found that all complexes adopt broad ranges of configurations, indicating high flexibility. We found unexpectedly that the DNA binding protein Nhp6 also binds to the C-terminal tails of FACT subunits, inducing more open geometries of FACT even in the absence of nucleosomes. Nhp6 therefore supports nucleosome unfolding by altering both the structure of FACT and the properties of nucleosomes. Complexes formed with FACT, Nhp6, and nucleosomes also produced a broad range of structures, revealing a large number of potential intermediates along a proposed unfolding pathway. The data suggest that Nhp6 has multiple roles before and during nucleosome unfolding by FACT, and that the process proceeds through a series of energetically similar intermediate structures, ultimately leading to an extensively unfolded form.
KW - Adenosine Triphosphate/chemistry
KW - DNA-Binding Proteins/chemistry
KW - High Mobility Group Proteins/chemistry
KW - Humans
KW - Microscopy, Electron, Transmission
KW - Nucleosomes/chemistry
KW - Protein Folding
KW - Saccharomyces cerevisiae Proteins/chemistry
KW - Saccharomyces cerevisiae/genetics
KW - Transcriptional Elongation Factors/chemistry
UR - http://www.scopus.com/inward/record.url?scp=85122651740&partnerID=8YFLogxK
U2 - 10.1038/s42003-021-02948-8
DO - 10.1038/s42003-021-02948-8
M3 - Article
C2 - 35013515
AN - SCOPUS:85122651740
SN - 2399-3642
VL - 5
SP - 2
JO - Communications Biology
JF - Communications Biology
IS - 1
M1 - 2
ER -