Stabilization of Nucleosomes by Histone Tails and by FACT Revealed by spFRET Microscopy
AbstractA correct chromatin structure is important for cell viability and is tightly regulated by numerous factors. Human protein complex FACT (facilitates chromatin transcription) is an essential factor involved in chromatin transcription and cancer development. Here FACT-dependent changes in the structure of single nucleosomes were studied with single-particle Förster resonance energy transfer (spFRET) microscopy using nucleosomes labeled with a donor-acceptor pair of fluorophores, which were attached to the adjacent gyres of DNA near the contact between H2A-H2B dimers. Human FACT and its version without the C-terminal domain (CTD) and the high mobility group (HMG) domain of the structure-specific recognition protein 1 (SSRP1) subunit did not change the structure of the nucleosomes, while FACT without the acidic C-terminal domains of the suppressor of Ty 16 (Spt16) and the SSRP1 subunits caused nucleosome aggregation. Proteolytic removal of histone tails significantly disturbed the nucleosome structure, inducing partial unwrapping of nucleosomal DNA. Human FACT reduced DNA unwrapping and stabilized the structure of tailless nucleosomes. CTD and/or HMG domains of SSRP1 are required for this FACT activity. In contrast, previously it has been shown that yeast FACT unfolds (reorganizes) nucleosomes using the CTD domain of SSRP1-like Pol I-binding protein 3 subunit (Pob3). Thus, yeast and human FACT complexes likely utilize the same domains for nucleosome reorganization and stabilization, respectively, and these processes are mechanistically similar. View Full-Text
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Valieva, M.E.; Gerasimova, N.S.; Kudryashova, K.S.; Kozlova, A.L.; Kirpichnikov, M.P.; Hu, Q.; Botuyan, M.V.; Mer, G.; Feofanov, A.V.; Studitsky, V.M. Stabilization of Nucleosomes by Histone Tails and by FACT Revealed by spFRET Microscopy. Cancers 2017, 9, 3.
Valieva ME, Gerasimova NS, Kudryashova KS, Kozlova AL, Kirpichnikov MP, Hu Q, Botuyan MV, Mer G, Feofanov AV, Studitsky VM. Stabilization of Nucleosomes by Histone Tails and by FACT Revealed by spFRET Microscopy. Cancers. 2017; 9(1):3.Chicago/Turabian Style
Valieva, Maria E.; Gerasimova, Nadezhda S.; Kudryashova, Kseniya S.; Kozlova, Anastasia L.; Kirpichnikov, Mikhail P.; Hu, Qi; Botuyan, Maria V.; Mer, Georges; Feofanov, Alexey V.; Studitsky, Vasily M. 2017. "Stabilization of Nucleosomes by Histone Tails and by FACT Revealed by spFRET Microscopy." Cancers 9, no. 1: 3.