Abstract
Genetic models suggested that SMARCA5 was required for DNA-templated events including transcription, DNA replication, and DNA repair. We engineered a degron tag into the endogenous alleles of SMARCA5, a catalytic component of the imitation switch complexes in three different human cell lines to define the effects of rapid degradation of this key regulator. Degradation of SMARCA5 was associated with a rapid increase in global nucleosome repeat length, which may allow greater chromatin compaction. However, there were few changes in nascent transcription within the first 6 h of degradation. Nevertheless, we demonstrated a requirement for SMARCA5 to control nucleosome repeat length at G1/S and during the S phase. SMARCA5 co-localized with CTCF and H2A.Z, and we found a rapid loss of CTCF DNA binding and disruption of nucleosomal phasing around CTCF binding sites. This spatiotemporal analysis indicates that SMARCA5 is continuously required for maintaining nucleosomal spacing.
Original language | English (US) |
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Pages (from-to) | 507-522.e6 |
Journal | Molecular Cell |
Volume | 83 |
Issue number | 4 |
DOIs | |
State | Published - Feb 16 2023 |
Keywords
- ATAC-seq
- CTCF
- H2A.Z
- MNase-seq
- PRO-seq
- SMARCA5
- chromatin
- nucleosome repeat length
ASJC Scopus subject areas
- Molecular Biology
- Cell Biology