The Cac2 subunit is essential for productive histone binding and nucleosome assembly in CAF-1

Year: 2017

Type: article

Abstract: Nucleosome assembly following DNA replication controls epigenome maintenance and genome integrity. Chromatin assembly factor 1 (CAF-1) is the histone chaperone responsible for histone (H3-H4)2 deposition following DNA synthesis. Structural and functional details for this chaperone complex and its interaction with histones are slowly emerging. Using hydrogen-deuterium exchange coupled to mass spectrometry, combined with in vitro and in vivo mutagenesis studies, we identified the regions involved in the direct interaction between the yeast CAF-1 subunits, and mapped the CAF-1 domains responsible for H3-H4 binding. The large subunit, Cac1 organizes the assembly of CAF-1. Strikingly, H3-H4 binding is mediated by a composite interface, shaped by Cac1-bound Cac2 and the Cac1 acidic region. Cac2 is indispensable for productive histone binding, while deletion of Cac3 has only moderate effects on H3-H4 binding and nucleosome assembly. These results define direct structural roles for yeast CAF-1 subunits and uncover a previously unknown critical function of the middle subunit in CAF-1.

Source: Scientific Reports

DOI: 10.1038/srep46274

Authors: Francesca Mattiroli, Yajie Gu, Jeremy L. Balsbaugh, Natalie G. Ahn, Karolin Luger

Institutions: Howard Hughes Medical Institute, University of Colorado Boulder, Colorado State University

Language: English

Topic: Genomics and Chromatin Dynamics

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: gold

Estimated APC paid: $2,690

Funders: Howard Hughes Medical Institute, U.S. Department of Energy, Colorado State University, KWF Kankerbestrijding, National Institutes of Health, Office of Science, University of Colorado Boulder, National Institute of General Medical Sciences, National Center for Research Resources, Argonne National Laboratory

OpenAlex ID: W2605910596