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.
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Gene/Complex | Qualifier | Gene Ontology Term | Annotation Extension | Evidence | Source | Assigned On |
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ARP5 | enables | enzyme regulator activity | has input Ino80 complex | IDA | SGD | 2021-01-13 |
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Evidence ID | Analyze ID | Interactor | Interactor Systematic Name | Interactor | Interactor Systematic Name | Allele | Assay | Annotation | Action | Phenotype | SGA score | P-value | Source | Reference | Note |
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Interactor | Interactor | Assay | Annotation | Action | Modification | |
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ACT1 | INO80 | Affinity Capture-Western | manually curated | Hit-Bait | No Modification | |
ACT1 | ARP5 | Affinity Capture-Western | manually curated | Hit-Bait | No Modification | |
ARP4 | ARP5 | Affinity Capture-Western | manually curated | Hit-Bait | No Modification | |
ARP5 | ARP8 | Affinity Capture-Western | manually curated | Bait-Hit | No Modification | |
ARP5 | IES2 | Affinity Capture-Western | manually curated | Bait-Hit | No Modification | |
ARP5 | RVB2 | Affinity Capture-Western | manually curated | Bait-Hit | No Modification | |
IES1 | INO80 | Affinity Capture-Western | manually curated | Hit-Bait | No Modification | |
IES3 | ARP5 | Affinity Capture-Western | manually curated | Hit-Bait | No Modification | |
IES5 | INO80 | Affinity Capture-Western | manually curated | Hit-Bait | No Modification | |
IES6 | INO80 | Affinity Capture-Western | manually curated | Hit-Bait | No Modification |