Structural visualization of key steps in nucleosome reorganization by human FACT
Abstract 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 domai...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2019-07-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-019-46617-7 |
| _version_ | 1818998900196900864 |
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| author | Kouta Mayanagi Kazumi Saikusa Naoyuki Miyazaki Satoko Akashi Kenji Iwasaki Yoshifumi Nishimura Kosuke Morikawa Yasuo Tsunaka |
| author_facet | Kouta Mayanagi Kazumi Saikusa Naoyuki Miyazaki Satoko Akashi Kenji Iwasaki Yoshifumi Nishimura Kosuke Morikawa Yasuo Tsunaka |
| author_sort | Kouta Mayanagi |
| collection | DOAJ |
| description | Abstract 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. |
| first_indexed | 2024-12-20T22:08:52Z |
| format | Article |
| id | doaj.art-9efebd6a9acf4ca9b718e8a5caa37b29 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-20T22:08:52Z |
| publishDate | 2019-07-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-9efebd6a9acf4ca9b718e8a5caa37b292022-12-21T19:25:13ZengNature PortfolioScientific Reports2045-23222019-07-019111410.1038/s41598-019-46617-7Structural visualization of key steps in nucleosome reorganization by human FACTKouta Mayanagi0Kazumi Saikusa1Naoyuki Miyazaki2Satoko Akashi3Kenji Iwasaki4Yoshifumi Nishimura5Kosuke Morikawa6Yasuo Tsunaka7Medical Institute of Bioregulation, Kyushu University, 3-1-1 Maidashi, Higashi-kuGraduate School of Science, Hiroshima University, 1-3-1 Kagamiyama, Higashi-HiroshimaLaboratory of Protein Synthesis and Expression, Institute for Protein Research, Osaka University, 3-2 YamadaokaGraduate School of Medical Life Science, Yokohama City University, 1-7-29 Suehiro-choLaboratory of Protein Synthesis and Expression, Institute for Protein Research, Osaka University, 3-2 YamadaokaGraduate School of Medical Life Science, Yokohama City University, 1-7-29 Suehiro-choDepartment of Gene Mechanisms, Graduate School of Biostudies, Kyoto University, Yoshida-konoemachiGraduate School of Medical Life Science, Yokohama City University, 1-7-29 Suehiro-choAbstract 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.https://doi.org/10.1038/s41598-019-46617-7 |
| spellingShingle | Kouta Mayanagi Kazumi Saikusa Naoyuki Miyazaki Satoko Akashi Kenji Iwasaki Yoshifumi Nishimura Kosuke Morikawa Yasuo Tsunaka Structural visualization of key steps in nucleosome reorganization by human FACT Scientific Reports |
| title | Structural visualization of key steps in nucleosome reorganization by human FACT |
| title_full | Structural visualization of key steps in nucleosome reorganization by human FACT |
| title_fullStr | Structural visualization of key steps in nucleosome reorganization by human FACT |
| title_full_unstemmed | Structural visualization of key steps in nucleosome reorganization by human FACT |
| title_short | Structural visualization of key steps in nucleosome reorganization by human FACT |
| title_sort | structural visualization of key steps in nucleosome reorganization by human fact |
| url | https://doi.org/10.1038/s41598-019-46617-7 |
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