Dataset Information

DBRWD3 manipulates chromatin signature by ubiquitination to regulate ORC

ABSTRACT: Ubiquitination and epigenetic modification both play essential roles in regulating cell fates and development, but little is known how two pathways crosstalk. A known chromatin-binding protein BRWD3, Bromodomain repeats and WD40 repeats domain contain protein 3, has been shown involved both in ubiquitination and histone modification regulation. BRWD3 has roles in multiple chromatin-related processes, including transcription, DNA replication, and chromatin modification. However, how ubiquitination is involved and what’s BRWD3’s ubiquitination targets are barely known. Here, we study the single Drosophila homolog dBRWD3 to uncover its mechanism. To find dBRWD3’s ubiquitination targets, we performed both dBRWD3 IP-MS and BRWD3 dependent ubiquitination IP-MS. Expectedly, the whole BRWD3-Cul4-DDB1 E3 ubiquitination complex and multiple histones are significantly enriched in dBRWD3-IP. Interestingly, multiple histone modification writers and the DNA replication initiation factor origin recognition complex (ORC) are among the intersection of two MS. ORC’s function is highly dependent on chromatin signature. dBRWD3 could provide a perspective on how chromatin signature is regulated by ubiquitination to regulate ORC function. In support of this, about 35% of ORC2 binding sites overlap with dBRWD3 binding sites among the genome. The H3K27ac occupancy rate of dBRWD3 related ORC sites is about 50% higher than that of dBRWD3 irrelevant ORC sites. Depletion of dBRWD3 significantly reduces DNA proliferation in the S phase. Further study will focus on determining which histone modification writer is targeted for ubiquitination and how does it contribute to the chromatin signature and ORC function.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Drosophila Melanogaster (fruit Fly)

TISSUE(S): Embryo

SUBMITTER: Jonathan Davies

LAB HEAD: Jared Nordman

PROVIDER: PXD039708 | Pride | 2023-10-24

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	20211020-JD-DH-002-IP-run2-150MIT-TMT11-0C_1.raw	Raw
	20211020-JD-DH-002-IP-run2-150MIT-TMT11-100C_7.raw	Raw
	20211020-JD-DH-002-IP-run2-150MIT-TMT11-10C_2.raw	Raw
	20211020-JD-DH-002-IP-run2-150MIT-TMT11-20C_3.raw	Raw
	20211020-JD-DH-002-IP-run2-150MIT-TMT11-40C_4.raw	Raw

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Publications

BRWD3 promotes KDM5 degradation to maintain H3K4 methylation levels.

Han Dongsheng D Schaffner Samantha H SH Davies Jonathan P JP Benton Mary Lauren ML Plate Lars L Nordman Jared T JT

Proceedings of the National Academy of Sciences of the United States of America 20230918 39

Histone modifications are critical for regulating chromatin structure and gene expression. Dysregulation of histone modifications likely contributes to disease states and cancer. Depletion of the chromatin-binding protein BRWD3 (Bromodomain and WD repeat-containing protein 3), a known substrate-specificity factor of the Cul4-DDB1 E3 ubiquitin ligase complex, results in increased H3K4me1 (H3 lysine 4 monomethylation) levels. The underlying mechanism linking BRWD3 and H3K4 methylation, however, ha ...[more]

PMID: 37722046

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