title: Assembly of the Arp5 (Actin-related Protein) Subunit Involved in Distinct INO80 Chromatin Remodeling Activities.
abstract: ATP-dependent chromatin remodeling, which repositions and restructures nucleosomes, is essential to all DNA-templated processes. The INO80 chromatin remodeling complex is an evolutionarily conserved complex involved in diverse cellular processes, including transcription, DNA repair, and replication. The functional diversity of the INO80 complex can, in part, be attributed to specialized activities of distinct subunits that compose the complex. Furthermore, structural analyses have identified biochemically discrete subunit modules that assemble along the Ino80 ATPase scaffold. Of particular interest is the Saccharomyces cerevisiae Arp5-Ies6 module located proximal to the Ino80 ATPase and the Rvb1-Rvb2 helicase module needed for INO80-mediated in vitro activity. In this study we demonstrate that the previously uncharacterized Ies2 subunit is required for Arp5-Ies6 association with the catalytic components of the INO80 complex. In addition, Arp5-Ies6 module assembly with the INO80 complex is dependent on distinct conserved domains within Arp5, Ies6, and Ino80, including the spacer region within the Ino80 ATPase domain. Arp5-Ies6 interacts with chromatin via assembly with the INO80 complex, as IES2 and INO80 deletion results in loss of Arp5-Ies6 chromatin association. Interestingly, ectopic addition of the wild-type Arp5-Ies6 module stimulates INO80-mediated ATP hydrolysis and nucleosome sliding in vitro. However, the addition of mutant Arp5 lacking unique insertion domains facilitates ATP hydrolysis in the absence of nucleosome sliding. Collectively, these results define the requirements of Arp5-Ies6 assembly, which are needed to couple ATP hydrolysis to productive nucleosome movement.
authors: From the Department of Biology, Stanford University, Stanford California 94305 and.
authors: Wei
authors: W
authors: Yao
authors: From the Department of Biology, Stanford University, Stanford California 94305 and.
authors: Sean L
authors: SL
authors: Beckwith
authors: From the Department of Biology, Stanford University, Stanford California 94305 and.
authors: Tina
authors: T
authors: Zheng
authors: From the Department of Biology, Stanford University, Stanford California 94305 and.
authors: Thomas
authors: T
authors: Young
authors: From the Department of Biology, Stanford University, Stanford California 94305 and.
authors: Van T
authors: VT
authors: Dinh
authors: Laboratory of Biochemistry and Molecular Biology, NCI, National Institutes of Health, Bethesda, Maryland 20892.
authors: Anand
authors: A
authors: Ranjan
authors: From the Department of Biology, Stanford University, Stanford California 94305 and ashbym@stanford.edu.
authors: Ashby J
authors: AJ
authors: Morrison
mesh_terms: D042002:Chromatin Assembly and Disassembly; D017434:Protein Structure, Tertiary; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000378:metabolism*
chemical_list: C483404:INO80 complex, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins
keywords: Arp5; INO80; Ies2; Ies6; chromatin; chromatin regulation; chromatin remodeling; histone; nucleosome; protein complex