TY - JOUR
T1 - Structural visualization of key steps in nucleosome reorganization by human FACT
AU - Mayanagi, Kouta
AU - Saikusa, Kazumi
AU - Miyazaki, Naoyuki
AU - Akashi, Satoko
AU - Iwasaki, Kenji
AU - Nishimura, Yoshifumi
AU - Morikawa, Kosuke
AU - Tsunaka, Yasuo
N1 - Funding Information:
We thank Dr. Yoshie Fujiwara (Kyoto University) for assistance. This work was supported in part by a Grant-in-Aid for Scientific Research (A) [JSPS KAKENHI Grant No. JP26251008 (to K.Mo.)], a Grant-in-Aid for Young Scientists (B) [JSPS KAKENHI Grant No. JP16K18528 (to K.S.)], a Grant-in-Aid for Scientific Research on Innovative Areas [JSPS KAKENHI Grant No. JP16H01410 (to K.Ma.)], and Grants-in-Aid for Scientific Research (C) [JSPS KAKENHI Grant Nos JP18K06089 (to K.Ma.), JP17K07313 (to S.A.) and JP18K06064 (to Y.T.)]. This work was partly supported by JST PREST grants [to K.Ma. (Grant No. JPMJPR12L9) and Y.T.] and Grants-in-Aid for Platform Project for Supporting Drug Discovery and Life Science Research (Basis for Supporting Innovative Drug Discovery and Life Science Research [BINDS]) from the Japan Agency for Medical Research and Development (AMED) [Japan under grant nos. JP17am0101076 (to S.A.), JP17am0101072j0001 (to K.I.), and JP17am0101073 (to Y.N.)]. This work was partly performed in the collaborative Research Project Program of the Medical Institute of Bioregulation, Kyushu University.
Publisher Copyright:
© 2019, The Author(s).
PY - 2019/12/1
Y1 - 2019/12/1
N2 - Facilitates chromatin transcription (FACT) is a histone chaperone, which accomplishes both nucleosome assembly and disassembly. Our combined cryo-electron microscopy (EM) and native mass spectrometry (MS) studies revealed novel key steps of nucleosome reorganization conducted by a Mid domain and its adjacent acidic AID segment of human FACT. We determined three cryo-EM structures of respective octasomes complexed with the Mid-AID and AID regions, and a hexasome alone. We discovered extensive contacts between a FACT region and histones H2A, H2B, and H3, suggesting that FACT is competent to direct functional replacement of a nucleosomal DNA end by its phosphorylated AID segment (pAID). Mutational assays revealed that the aromatic and phosphorylated residues within pAID are essential for octasome binding. The EM structure of the hexasome, generated by the addition of Mid-pAID or pAID, indicated that the dissociation of H2A-H2B dimer causes significant alteration from the canonical path of the nucleosomal DNA.
AB - Facilitates chromatin transcription (FACT) is a histone chaperone, which accomplishes both nucleosome assembly and disassembly. Our combined cryo-electron microscopy (EM) and native mass spectrometry (MS) studies revealed novel key steps of nucleosome reorganization conducted by a Mid domain and its adjacent acidic AID segment of human FACT. We determined three cryo-EM structures of respective octasomes complexed with the Mid-AID and AID regions, and a hexasome alone. We discovered extensive contacts between a FACT region and histones H2A, H2B, and H3, suggesting that FACT is competent to direct functional replacement of a nucleosomal DNA end by its phosphorylated AID segment (pAID). Mutational assays revealed that the aromatic and phosphorylated residues within pAID are essential for octasome binding. The EM structure of the hexasome, generated by the addition of Mid-pAID or pAID, indicated that the dissociation of H2A-H2B dimer causes significant alteration from the canonical path of the nucleosomal DNA.
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U2 - 10.1038/s41598-019-46617-7
DO - 10.1038/s41598-019-46617-7
M3 - Article
C2 - 31308435
AN - SCOPUS:85069041844
SN - 2045-2322
VL - 9
JO - Scientific reports
JF - Scientific reports
IS - 1
M1 - 10183
ER -