FACT (facilitates chromatin transcription, [1] sometimes facilitates chromatin transactions [2] [3]) is a heterodimeric protein complex that affects eukaryotic RNA polymerase II (Pol II) transcription elongation both in vitro and in vivo. It was discovered in 1998 as a factor purified from human cells that was essential for productive, in vitro Pol II transcription on a chromatinized DNA template. [4]
FACT consists of 140 and 80 kilodalton (kDa) subunits. The 140 kDa subunit is encoded by the human gene SUPT16H, (SPT16 in S. cerevisiae) while the 80 kDa subunit is encoded by the human gene SSRP1 (POB3 in S. cerevisiae). Both of these subunits in yeast affect Pol II transcription elongation, and purified human FACT binds specifically to mononucleosomes and the histone H2A/ H2B dimer, but not to the H3/ H4 tetramer (see: Nucleosome core particle) or Pol II. [5]
Co-immunoprecipitation assays with tagged recombinant proteins showed that the Spt16 subunit interacts with H2A/H2B dimers and mononucleosomes, but not H3/H4 tetramers, whereas the SSRP1 subunit interacts only with H3/H4 tetramers and not mononucleosomes. Deletion of the highly acidic C-terminus of Spt16 (a common feature of known histone chaperones) does not prevent Spt16 from forming a stable complex with SSRP1, but it does eliminate interaction with mononucleosomes and ability to stimulate in vitro transcription on chromatinized templates. The two subunits together, but neither alone, can stimulate formation of nucleosomes from free histones and DNA (histone chaperone activity). These two subunits are highly conserved across all eukaryotes, and in addition to transcription, have been shown to affect DNA repair and replication as well. [6]
In cells, FACT is enriched on parts of the genome involved in actively elongating Pol II, as seen in fluorescent-antibody staining of Drosophila polytene chromosomes and chromatin immunoprecipitation (ChIP) assays on Drosophila Kc cell extracts. [7]
References
- ^ Winkler, Duane D.; Luger, Karolin (2011-05-27). "The Histone Chaperone FACT: Structural Insights and Mechanisms for Nucleosome Reorganization". The Journal of Biological Chemistry. 286 (21): 18369–18374. doi: 10.1074/jbc.R110.180778. ISSN 0021-9258. PMC 3099653. PMID 21454601.
- ^ Martin, Benjamin J. E.; Chruscicki, Adam T.; Howe, LeAnn J. (2018-11-01). "Transcription Promotes the Interaction of the FAcilitates Chromatin Transactions (FACT) Complex with Nucleosomes in Saccharomyces cerevisiae". Genetics. 210 (3): 869–881. doi: 10.1534/genetics.118.301349. ISSN 0016-6731. PMC 6218215. PMID 30237209.
- ^ Frost, Jennifer M.; Kim, M. Yvonne; Park, Guen Tae; Hsieh, Ping-Hung; Nakamura, Miyuki; Lin, Samuel J. H.; Yoo, Hyunjin; Choi, Jaemyung; Ikeda, Yoko; Kinoshita, Tetsu; Choi, Yeonhee (2018-05-15). "FACT complex is required for DNA demethylation at heterochromatin during reproduction in Arabidopsis". Proceedings of the National Academy of Sciences. 115 (20): E4720–E4729. doi: 10.1073/pnas.1713333115. ISSN 0027-8424. PMC 5960277. PMID 29712855.
- ^ Orphanides, George; LeRoy, Gary; Chang, Chun-Hsiang; Luse, Donal S.; Reinberg, Danny (1998). "FACT, a Factor that Facilitates Transcript Elongation through Nucleosomes". Cell. 92 (1): 105–116. doi: 10.1016/S0092-8674(00)80903-4. PMID 9489704. S2CID 11248520.
- ^ Orphanides, George; Wu, Wei-Hua; Lane, William S.; Hampsey, Michael; Reinberg, Danny (1999). "The chromatin-specific transcription elongation factor FACT comprises human SPT16 and SSRP1 proteins". Nature. 400 (6741): 284–288. Bibcode: 1999Natur.400..284O. doi: 10.1038/22350. PMID 10421373. S2CID 4300397.
- ^ Belotserkovskaya, Rimma; Oh, Sangtaek; Bondarenko, Vladimir A.; Orphanides, George; Studitsky, Vasily; Reinberg, Danny (2003). "FACT facilitates transcription-dependent nucleosome alteration". Science. 301 (5636): 1090–1093. Bibcode: 2003Sci...301.1090B. doi: 10.1126/science.1085703. PMID 12934006. S2CID 26667338.
- ^ Saunders, Abbie; Werner, Janis; Andrulis, Erik D.; Nakayama, Takahiro; Hirose, Susumu; Reinberg, Danny; Lis, John T. (2003). "Tracking FACT and the RNA Polymerase II Elongation Complex Through Chromatin in Vivo". Science. 301 (5636): 1094–1096. Bibcode: 2003Sci...301.1094S. doi: 10.1126/science.1085712. PMID 12934007. S2CID 10064786.
Further reading
Reinberg, D., Sims, R. (2006). de FACTo nucleosome dynamics. Journal of Biological Chemistry, 281(33) 23297-23391.