| 2018 |
Cryo-EM structure of the evolutionarily conserved INO80 core complex from Chaetomium thermophilum bound to a nucleosome at 4.3 Å (major parts at 3.7 Å) revealed that Arp5 and Ies6 (INO80C) bind superhelical locations -2 and -3 to act as a counter grip for the ATPase motor on the opposite side of the H2A-H2B dimer. The Arp5 insertion domain forms a grappler element that contacts the nucleosome dyad and packs against histone H2A-H2B near the acidic patch, while the ATPase motor pumps entry DNA into the nucleosome against this Arp5-Ies6 grip. |
Cryo-EM structure determination; biochemical assays |
Nature |
High |
29643509
|
| 2018 |
Cryo-EM structure of the human INO80 complex bound to a nucleosome showed that the ARP5-IES6 (INO80C) module makes contacts on the opposite face of the nucleosome from the motor domains, and that histone H3 tails (rather than H4 tails as in other remodelers) regulate INO80 motor domain activity in a manner dependent on this ARP5-IES6 arrangement. |
Cryo-EM structure determination; biochemical regulation assays |
Nature |
High |
29643506
|
| 2011 |
The human INO80 complex is organized into three modules assembling on distinct domains of hIno80 ATPase. The third module, comprising the hIno80 Snf2 ATPase domain, Ies2, Ies6 (INO80C), Tip49a, Tip49b, and Arp5, is part of the evolutionarily conserved core required for ATP-dependent nucleosome remodeling activity. |
Subcomplex purification; biochemical reconstitution; ATPase and nucleosome remodeling assays |
The Journal of biological chemistry |
High |
21303910
|
| 2013 |
In the human INO80 complex, Ies6 (INO80C) and Arp5 function together to promote binding of the Ino80 ATPase to nucleosomes, whereas Ies2 functions as a potent activator of the intrinsic ATPase catalytic activity. Ies6 and Arp5 thus regulate substrate recognition rather than intrinsic catalysis. |
Biochemical reconstitution; ATPase assays; nucleosome-binding assays with purified subcomplexes |
Proceedings of the National Academy of Sciences of the United States of America |
High |
24297934
|
| 2015 |
The Ies6/Arp5 module is essential for INO80 complex chromatin remodeling activity in vitro, and controls conformational changes that couple nucleosome binding to remodeling. By contrast, the Arp8/Arp4/Act1 module enhances nucleosome-binding affinity but is largely dispensable for remodeling. EM class averages and mass spectrometry positioned these modules within the overall architecture. |
Electron microscopy; mass spectrometry; biochemical remodeling assays with defined subunit deletions |
Nature communications |
High |
25964121
|
| 2016 |
Arp5 and Ies6 (INO80C) form an abundant and distinct subcomplex in vivo in yeast, stimulate INO80-mediated ATP hydrolysis and nucleosome sliding in vitro, and their genomic occupancy correlates with nucleosome positioning at transcriptional start sites and expression levels of >1,000 INO80-regulated genes enriched in energy metabolism pathways. Loss of ies6 leads to decreased glycolytic gene expression and elevated mitochondrial potential. |
Co-immunoprecipitation; in vitro ATPase and nucleosome sliding assays; genomic ChIP-seq; gene expression analysis |
Molecular and cellular biology |
High |
26755556
|
| 2015 |
Assembly of the Arp5-Ies6 module with the INO80 complex requires distinct conserved domains within Arp5, Ies6, and the Ino80 spacer region. Ies2 is required for Arp5-Ies6 association with the catalytic components; loss of IES2 or INO80 abolishes Arp5-Ies6 chromatin association. A mutant Arp5 lacking its insertion domains can stimulate ATPase activity but not nucleosome sliding, indicating the insertion domain couples hydrolysis to translocation. |
Domain deletion mutagenesis; Co-immunoprecipitation; chromatin immunoprecipitation; in vitro ATPase and nucleosome sliding assays |
The Journal of biological chemistry |
High |
26306040
|
| 2016 |
Using a recombinant minimal human INO80 core complex, Arp5/Ies6 and Ies2 were shown to regulate nucleosome sliding synergistically and antagonistically. Inositol hexaphosphate (IP6) is a non-competitive inhibitor that blocks the stimulatory effect of nucleosomes on ATPase activity, with its binding site in the C-terminal region of Ino80. An Arp5 bypass mutation restores activity in the absence of Ies2, revealing coupling between Ies2 and Arp5/Ies6 in controlling ATP hydrolysis-to-sliding coupling. |
Recombinant protein reconstitution; ATPase assays; nucleosome sliding assays; inhibitor studies; mutagenesis |
Nucleic acids research |
High |
27257055
|
| 2012 |
Loss of the Ies6 subunit (INO80C) in budding yeast phenocopies loss of the Ino80 catalytic subunit, causing rapid polyploidy, defective chromosome segregation, and altered pericentric chromatin structure due to misincorporation of H2A.Z into pericentric nucleosomes. Ies6 is thus critical for INO80 function in preventing H2A.Z misincorporation at centromeres. |
Genetic deletion; flow cytometry (ploidy measurement); chromatin immunoprecipitation; live-cell imaging of chromosome segregation |
Genes & development |
High |
23207916
|
| 2009 |
In fission yeast, deletion of ies6 causes defects in DNA damage repair, response to replication stress, and nucleotide metabolism, phenocopying deletion of other INO80 complex subunits. The Ino80 complex from fission yeast containing Ies6 mediates ATP-dependent nucleosome remodeling in vitro. |
Genetic deletion; DNA damage sensitivity assays; in vitro nucleosome remodeling assays |
Molecular and cellular biology |
Medium |
19933844
|
| 2018 |
In yeast, the Ies6 subunit module displays a divergent genetic interaction signature that links the INO80 complex to metabolic homeostasis. ies6 mutants show disrupted mitochondrial maintenance, and INO80 (including its Ies6 module) is needed for TORC1-mediated signaling to chromatin, as ino80 mutants exhibit defective transcriptional profiles and altered histone acetylation of TORC1-responsive genes. |
Genetic interaction screen; gene expression analysis; histone modification assays |
PLoS genetics |
Medium |
29462149
|
| 2022 |
ACTR5 (Arp5) and its interacting partner IES6 (INO80C) show a distinct, HCC-specific functional signature compared to other INO80 complex members in CRISPR tiling scans, suggesting an INO80-independent mechanism of ACTR5/IES6 in supporting hepatocellular carcinoma cell proliferation. Suppression of ACTR5 activated CDKN2A and ablated CDK/E2F-driven cell cycle signaling. |
CRISPR interference screen; CRISPR gene tiling scans; gene expression analysis; xenograft tumor assays |
Science advances |
Medium |
36563143
|
| 2017 |
INO80 complex function in homologous recombination includes at least two distinct steps: DNA end resection and presynaptic filament formation. The second function is linked to H2A.Z: in the absence of H2A.Z, presynaptic filament formation and HR are restored in INO80-C-deficient yeast mutants, indicating that INO80-C facilitates HR by removing H2A.Z to promote filament formation. |
Genetic epistasis (double mutant analysis); fluorescence microscopy of repair intermediates; HR frequency assays in yeast |
Cell reports |
Medium |
28514650
|
| 2016 |
In budding yeast, the INO80 complex cooperates with Mec1 (ATR) and PAF1C to remove RNAPII from transcribed genes near early-firing replication origins upon replication stress (hydroxyurea). This removal is required for efficient replication fork restart; failure to evict RNAPII in ino80 mutants correlates with inability to restart stalled forks. |
Genetic epistasis; ChIP; proteomic analyses; replication fork restart assays |
Genes & development |
Medium |
26798134
|
| 2020 |
INO80 complex co-localizes with the origin recognition complex (ORC) at yeast replication origins and replication initiation sites in mouse ESCs, preventing pervasive transcription through origin sequences. Genetic studies show that INO80C and Mot1/NC2 function through distinct pathways to limit origin transcription; absence of INO80C leads to formation of new DNA double-strand breaks at origins. |
ChIP-seq; nascent transcript sequencing; genetic epistasis; DSB detection assays |
Cell reports |
Medium |
32905765
|
| 2021 |
During DNA damage in yeast, INO80C-dependent recruitment of five ubiquitin-conjugating factors (Rad6, Bre1, Pep5, Ufd4, and Rsp5) contributes to core and linker histone depletion at damaged chromatin, reducing chromatin compaction and enhancing DNA locus mobility and strand invasion kinetics during homology-driven repair. |
Chromatin-associated proteomics; genetic epistasis; live-cell imaging; strand invasion assays |
Molecular cell |
Medium |
33529595
|
| 2023 |
Abasic sites and UV-irradiation damage abolish the DNA translocation activity of INO80-C by compromising ATP hydrolysis within the Ino80 catalytic subunit, while nucleosome binding remains unaffected. INO80-C also facilitates cleavage of abasic (AP) sites by AP-endonuclease 1 (APE1) independently of its DNA translocation activity. |
In vitro DNA translocation assays; ATPase assays with damaged substrates; AP site cleavage assay; nucleosome binding assays |
The Journal of biological chemistry |
Medium |
37696438
|
| 2024 |
Loss of cytoplasmic actin filaments (via TORC2 inhibition or Las17 degradation) raises nuclear actin levels, which in complex with Arp4 is an essential subunit of INO80C. Genetic ablation of INO80C activity leads to partial resistance to yeast chromosome shattering (YCS), suggesting that elevated nuclear G-actin stimulates INO80C to increase DNA polymerase processivity and thereby converts single-strand lesions into double-strand breaks. |
Phosphoproteomics; auxin-induced protein degradation; genetic ablation; nuclear actin quantification |
Nature communications |
Medium |
39548059
|
| 2026 |
Deletion of IES6 in yeast reduces genome-wide nucleosome spacing by 3 bp and disrupts regular nucleosome arrays across most genes. IES6 deletion is synthetically lethal with deletion of ISW2 (an ISWI-family remodeler), indicating functional redundancy in nucleosome organization. INO80 binding directly predicts the role of Ies6 in nucleosome organization, whereas gene expression changes do not correlate with altered spacing. |
Genome-wide nucleosome mapping (MNase-seq); genetic epistasis (synthetic lethality); ChIP-seq |
Scientific reports |
Medium |
41720950
|
| 2018 |
In the human INO80 complex, biochemical approaches indicated that INO80-C and the H3K27 acetyltransferase P300 physically interact, suggesting they may jointly coordinate chromatin accessibility at canonical INO80 target sites. |
Co-immunoprecipitation; ChIP-seq for INO80 subunits |
G3 (Bethesda, Md.) |
Low |
29432129
|