SMT Datasets from: Jeronimo et al., 2021. "FACT is recruited to the +1 nucleosome of transcribed genes and spreads in a Chd1-dependent manner"
Description
Title: FACT is recruited to the +1 nucleosome of transcribed genes and spreads in a Chd1-dependent manner Célia Jeronimo1, Andrew Angel2, Vu Q. Nguyen3, Jee Min Kim3, Christian Poitras1, Elie Lambert1, Pierre Collin1, Jane Mellor2, Carl Wu3,4 and François Robert1,5,6* 1 Institut de recherches cliniques de Montréal, 110 Avenue des Pins Ouest, Montréal, QC H2W 1R7, Canada. 2Department of Biochemistry, University of Oxford, South Parks Road, Oxford, OX1 3QU, UK. 3Department of Biology, Johns Hopkins University, Baltimore, MD 21218, USA. 4Department of Molecular Biology and Genetics, Johns Hopkins School of Medicine, Baltimore, MD 21287, USA. 5 Département de Médecine, Faculté de Médecine, Université de Montréal, 2900 Boul. Édouard-Montpetit, Montréal, QC H3T 1J4, Canada. 6 Lead Contact *Correspondence: francois.robert@ircm.qc.ca Abstract: The histone chaperone FACT occupies transcribed regions where it plays prominent roles in maintaining chromatin integrity and preserving epigenetic information. How it is targeted to transcribed regions, however, remains unclear. Proposed models include docking on the RNA polymerase II (RNAPII) C-terminal domain (CTD), recruitment by elongation factors, recognition of modified histone tails and binding partially disassembled nucleosomes. Here, we systematically tested these and other scenarios in Saccharomyces cerevisiae and found that FACT binds transcribed chromatin, not RNAPII. Through a combination of high-resolution genome-wide mapping, single-molecule tracking and mathematical modeling, we propose that FACT recognizes the +1 nucleosome, as it is partially unwrapped by the engaging RNAPII, and spreads to downstream nucleosomes aided by the chromatin remodeler Chd1. Our work clarifies how FACT interacts with genes, suggests a processive mechanism for FACT function, and provides a framework to further dissect the molecular mechanisms of transcription-coupled histone chaperoning. File Descriptions: SMT datasets: Filenames include information about: strain number, HaloTagged factor name, condition, biological replicate number, laser power (mW), frame rate (ms), and movie number. .mat files are raw output files from tracking software DiaTrack 3.04, and contains trajectory information (X coordinate, Y coordinate, intensity, goodness of fit, etc.) from each movie. .tif files are "MASK" files where the nuclear regions was masked based on the maximum intensity Z-projection of selected substacks of each movie to filter out trajectories found outside of the nucleus in the subsequent analysis steps. Contact information: Name: Carl Wu Organization: Johns Hopkins University Email: wuc@jhu.edu