FCCC LOGO Faculty Publications
McCullough LL , Connell Z , Xin H , Studitsky VM , Feofanov AV , Valieva ME , Formosa T
Functional roles of the DNA-binding HMGB domain in the histone chaperone FACT in nucleosome reorganization
J Biol Chem. 2018 Apr 20;293(16) :6121-6133
PMID: 29514976    PMCID: PMC5912460    URL: https://www.ncbi.nlm.nih.gov/pubmed/29514976
Back to previous list
The essential histone chaperone FAcilitates Chromatin Transcription (FACT) promotes both nucleosome assembly and disassembly. FACT is a heterodimer of Spt16 with either SSRP1 or Pob3, differing primarily by the presence of a high-mobility group B (HMGB) DNA-binding domain furnished only by SSRP1. Yeast FACT lacks the intrinsic HMGB domain found in SSRP1-based homologs such as human FACT, but yeast FACT activity is supported by Nhp6, which is a freestanding, single HMGB domain protein. The importance of histone binding by FACT domains has been established, but the roles of DNA binding activity remain poorly understood. Here, we examined these roles by fusing single or multiple HMGB modules to Pob3 to mimic SSRP1 or to test the effects of extended DNA-binding capacity. Human FACT and a yeast mimic both required Nhp6 to support nucleosome reorganization in vitro, indicating that a single intrinsic DNA-binding HMGB module is insufficient for full FACT activity. Three fused HMGB modules supported activity without Nhp6 assistance, but this FACT variant did not efficiently release from nucleosomes and was toxic in vivo. Notably, intrinsic DNA-binding HMGB modules reduced the DNA accessibility and histone H2A-H2B dimer loss normally associated with nucleosome reorganization. We propose that DNA bending by HMGB domains promotes nucleosome destabilization and reorganization by exposing FACT's histone binding sites, but DNA bending also produces DNA curvature needed to accommodate nucleosome assembly. Intrinsic DNA bending activity therefore favors nucleosome assembly by FACT over nucleosome reorganization, but excessive activity impairs FACT release, suggesting a quality control checkpoint during nucleosome assembly.
1083-351x McCullough, Laura L Connell, Zaily Xin, Hua Studitsky, Vasily M Feofanov, Alexey V Valieva, Maria E Formosa, Tim Journal Article United States J Biol Chem. 2018 Mar 7. pii: RA117.000199. doi: 10.1074/jbc.RA117.000199.