The Cac1 subunit of histone chaperone CAF-1 organizes CAF-1-H3/H4 architecture and tetramerizes histones.
The histone chaperone Chromatin Assembly Factor 1 (CAF-1) deposits tetrameric (H3/H4)2histones onto newly-synthesized DNA during DNA replication. To understand the mechanism of the tri-subunit CAF-1 complex in this process, we investigated the protein-protein interactions within the CAF-1-H3/H4 architecture using biophysical and biochemical approaches. Hydrogen/deuterium exchange and chemical cross-linking coupled to mass spectrometry reveal interactions that are essential for CAF-1 function in budding yeast, and importantly indicate that the Cac1 subunit functions as a scaffold within the CAF-1-H3/H4 complex. Cac1 alone not only binds H3/H4 with high affinity, but also promotes histone tetramerization independent of the other subunits. Moreover, we identify a minimal region in the C-terminus of Cac1, including the structured winged helix domain and glutamate/aspartate-rich domain, which is sufficient to induce (H3/H4)2tetramerization. These findings reveal a key role of Cac1 in histone tetramerization, providing a new model for CAF-1-H3/H4 architecture and function during eukaryotic replication.
Department of Pharmacology, University of Colorado School of Medicine, Aurora, United States.
Wallace H
WH
Liu
Department of Pharmacology, University of Colorado School of Medicine, Aurora, United States.
Sarah C
SC
Roemer
Department of Pharmacology, University of Colorado School of Medicine, Aurora, United States.
Yeyun
Y
Zhou
Department of Pathology and Laboratory Medicine, Weill Cornell Medicine, New York, United States.
Zih-Jie
ZJ
Shen
Department of Epigenetics and Molecular Carcinogenesis, MD Anderson Cancer Center, Houston, United States.
Briana K
BK
Dennehey
Department of Chemistry and Biochemistry, University of Colorado, Boulder, Boulder, United States.|BioFrontiers Institute, University of Colorado, Boulder, Boulder, United States.
Jeremy L
JL
Balsbaugh
Department of Chemistry and Biochemistry, University of Colorado, Boulder, Boulder, United States.|BioFrontiers Institute, University of Colorado, Boulder, Boulder, United States.
Jennifer C
JC
Liddle
Program in Structural Biology and Biochemistry, University of Colorado School of Medicine, Aurora, United States.
Travis
T
Nemkov
Department of Chemistry and Biochemistry, University of Colorado, Boulder, Boulder, United States.|BioFrontiers Institute, University of Colorado, Boulder, Boulder, United States.
Natalie G
NG
Ahn
Program in Structural Biology and Biochemistry, University of Colorado School of Medicine, Aurora, United States.
Kirk C
KC
Hansen
Department of Pathology and Laboratory Medicine, Weill Cornell Medicine, New York, United States.|Department of Epigenetics and Molecular Carcinogenesis, MD Anderson Cancer Center, Houston, United States.
Jessica K
http://orcid.org/0000-0001-9765-1659
JK
Tyler
Department of Pharmacology, University of Colorado School of Medicine, Aurora, United States.|Program in Structural Biology and Biochemistry, University of Colorado School of Medicine, Aurora, United States.
Mair Ea
http://orcid.org/0000-0003-0862-235X
ME
Churchill
Nucleosome; S. cerevisiae; biophysics; chromosomes; genes; histone chaperone; histones; mass spectrometry; protein-protein interactions; structural biology
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