摘要：SummaryThe FACT (facilitates chromatin transcription) complex, comprising SPT16 and SSRP1, conducts structural alterations during nucleosome unwrapping. Our previous cryoelectron microscopic (cryo-EM) analysis revealed the first intermediate structure of an unwrapped nucleosome with human FACT, in which 112-bp DNA and the phosphorylated intrinsically disordered (pAID) segment of SPT16 jointly wrapped around the histone core instead of 145-bp DNA. Using NMR, here we clarified that the histone H3 N-terminal tails, unobserved in the cryo-EM structure, adopt two different conformations reflecting their asymmetric locations at entry/exit sites: one corresponds to the original nucleosome site buried in two DNA gyres (DNA side), whereas the other, comprising pAID and DNA, is more exposed to the solvent (pAID side). NMR real-time monitoring showed that H3 acetylation is faster on the pAID side than on the DNA side. Our findings highlight that accessible conformations of H3 tails are created by the replacement of nucleosomal DNA with pAID.Graphical AbstractDisplay OmittedHighlights•H3 N-tail, restricted to two DNA gyres of nucleosome, is protected from Gcn5•H3 N-tail is dynamically exposed by replacement of nucleosomal DNA with pAID of FACT•Gcn5 efficiently acetylates accessible H3 N-tail of nucleosome with FACT•FACT acts as a modulator for dynamic behavior of H3 tails in nucleosomeBiochemistry; Molecular Biology; Structural Biology
