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1 Identification of replisome-associated proteins in Drosophila embryos and cultured cells using iPOND coupled to quantitative mass spectrometry

ABSTRACT: Replication of the eukaryotic genome requires the assembly of thousands of replisomes that must work in concert to accurately replicate a cell’s genetic and epigenetic information. Defining replisome-associated proteins is a key step in understanding how genomes are replicated and repaired in the context of chromatin to maintain genome stability. To identify replisome-associated proteins, we performed iPOND (Isolation of Proteins on Nascent DNA) coupled to quantitative mass spectrometry in Drosophila embryos and cultured cells. We identified 76 and 416 replisome-associated proteins in post-MZT embryos and Drosophila cultured S2 cells, respectively . By performing a targeted screen of a subset of these proteins, we demonstrate that BRWD3, a targeting specificity factor for the DDB1/Cul4 ubiquitin ligase complex (CRL4), functions at the replisome to promote replication fork progression and maintain genome stability. Altogether, our work provides a valuable resource for those interested in the DNA replication, repair and chromatin assembly during development.

INSTRUMENT(S): Orbitrap Exploris 480, Q Exactive HF

ORGANISM(S): Drosophila Melanogaster (fruit Fly)

TISSUE(S): Embryo, Late Embryonic Stage, Cell Culture

SUBMITTER: Madison Wright

LAB HEAD: Jared T. Nordman

PROVIDER: PXD031165 | Pride | 2022-01-27

REPOSITORIES: Pride

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