The SAGA histone acetyltransferase module targets SMC5/6 to specific genes.
Ada2
Chromatin accessibility
DNA repair
Gcn5
Gene regions
Genetic and protein–protein interactions
Histone H3K9ac acetylation
Nse3 KITE
SAGA histone acetyltransferase module
SMC5/6 complex targeting
rDNA
Journal
Epigenetics & chromatin
ISSN: 1756-8935
Titre abrégé: Epigenetics Chromatin
Pays: England
ID NLM: 101471619
Informations de publication
Date de publication:
16 02 2023
16 02 2023
Historique:
received:
22
10
2022
accepted:
02
02
2023
entrez:
16
2
2023
pubmed:
17
2
2023
medline:
18
2
2023
Statut:
epublish
Résumé
Structural Maintenance of Chromosomes (SMC) complexes are molecular machines driving chromatin organization at higher levels. In eukaryotes, three SMC complexes (cohesin, condensin and SMC5/6) play key roles in cohesion, condensation, replication, transcription and DNA repair. Their physical binding to DNA requires accessible chromatin. We performed a genetic screen in fission yeast to identify novel factors required for SMC5/6 binding to DNA. We identified 79 genes of which histone acetyltransferases (HATs) were the most represented. Genetic and phenotypic analyses suggested a particularly strong functional relationship between the SMC5/6 and SAGA complexes. Furthermore, several SMC5/6 subunits physically interacted with SAGA HAT module components Gcn5 and Ada2. As Gcn5-dependent acetylation facilitates the accessibility of chromatin to DNA-repair proteins, we first analysed the formation of DNA-damage-induced SMC5/6 foci in the Δgcn5 mutant. The SMC5/6 foci formed normally in Δgcn5, suggesting SAGA-independent SMC5/6 localization to DNA-damaged sites. Next, we used Nse4-FLAG chromatin-immunoprecipitation (ChIP-seq) analysis in unchallenged cells to assess SMC5/6 distribution. A significant portion of SMC5/6 accumulated within gene regions in wild-type cells, which was reduced in Δgcn5 and Δada2 mutants. The drop in SMC5/6 levels was also observed in gcn5-E191Q acetyltransferase-dead mutant. Our data show genetic and physical interactions between SMC5/6 and SAGA complexes. The ChIP-seq analysis suggests that SAGA HAT module targets SMC5/6 to specific gene regions and facilitates their accessibility for SMC5/6 loading.
Sections du résumé
BACKGROUND
Structural Maintenance of Chromosomes (SMC) complexes are molecular machines driving chromatin organization at higher levels. In eukaryotes, three SMC complexes (cohesin, condensin and SMC5/6) play key roles in cohesion, condensation, replication, transcription and DNA repair. Their physical binding to DNA requires accessible chromatin.
RESULTS
We performed a genetic screen in fission yeast to identify novel factors required for SMC5/6 binding to DNA. We identified 79 genes of which histone acetyltransferases (HATs) were the most represented. Genetic and phenotypic analyses suggested a particularly strong functional relationship between the SMC5/6 and SAGA complexes. Furthermore, several SMC5/6 subunits physically interacted with SAGA HAT module components Gcn5 and Ada2. As Gcn5-dependent acetylation facilitates the accessibility of chromatin to DNA-repair proteins, we first analysed the formation of DNA-damage-induced SMC5/6 foci in the Δgcn5 mutant. The SMC5/6 foci formed normally in Δgcn5, suggesting SAGA-independent SMC5/6 localization to DNA-damaged sites. Next, we used Nse4-FLAG chromatin-immunoprecipitation (ChIP-seq) analysis in unchallenged cells to assess SMC5/6 distribution. A significant portion of SMC5/6 accumulated within gene regions in wild-type cells, which was reduced in Δgcn5 and Δada2 mutants. The drop in SMC5/6 levels was also observed in gcn5-E191Q acetyltransferase-dead mutant.
CONCLUSION
Our data show genetic and physical interactions between SMC5/6 and SAGA complexes. The ChIP-seq analysis suggests that SAGA HAT module targets SMC5/6 to specific gene regions and facilitates their accessibility for SMC5/6 loading.
Identifiants
pubmed: 36793083
doi: 10.1186/s13072-023-00480-z
pii: 10.1186/s13072-023-00480-z
pmc: PMC9933293
doi:
Substances chimiques
Acetyltransferases
EC 2.3.1.-
Carrier Proteins
0
Cell Cycle Proteins
0
Chromatin
0
DNA
9007-49-2
Gcn5 protein, S pombe
EC 2.3.1.48
Histone Acetyltransferases
EC 2.3.1.48
Nse4 protein, S pombe
0
Schizosaccharomyces pombe Proteins
0
Smc5 protein, S pombe
0
smc6 protein, S pombe
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
6Subventions
Organisme : Ministerstvo Školství, Mládeže a Tělovýchovy
ID : LTC20033
Organisme : Masarykova Univerzita
ID : MUNI/R/1142/2021
Organisme : Univerzita Karlova v Praze
ID : PRIMUS/MED/26
Informations de copyright
© 2023. The Author(s).
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