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Replisome composition of stalled, terminating replication forks

ABSTRACT: Topological stress can cause replication forks to stall as they converge upon one another during termination of vertebrate DNA synthesis. However, replication forks ultimately overcome topological stress and complete DNA synthesis, suggesting that alternative mechanisms can overcome topological stress. We performed a proteomic analysis of converging replication forks that were stalled by topological stress induced by loss or inhibition of topoisomerase IIα (TOP2α). Plasmid DNA was replicated in mock- or TOP2α-depleted Xenopus egg extracts as previously described (Heintzman et al. 2019). In parallel, replication was performed in the presence of the TOP2 inhibitor ICRF-193 (‘TOP2-i’) as an alternate means of preventing TOP2 activity (Heintzman et al. 2019). Chromatinized plasmid DNA was recovered 18 minutes after the onset of DNA synthesis, when most forks have normally merged but are stalled when TOP2 activity is prevented (Heintzman et al. 2019). Chromatin-bound proteins were recovered (Larsen et al. 2019) then analyzed by chromatin mass spectrometry and quantified by label free quantification.

INSTRUMENT(S): Q Exactive Plus

ORGANISM(S): Xenopus Laevis (african Clawed Frog)

TISSUE(S): Egg Cell, Oocyte

SUBMITTER: Kristie Lindsey Rose

LAB HEAD: Kristie Lindsey Rose

PROVIDER: PXD031170 | Pride | 2023-02-21

REPOSITORIES: Pride

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			Action	DRS
	MaxquantLFQ_Proj5454_proteinGroups.txt	Txt
	Maxquant_Proj5454_peptides.txt	Txt
	checksum.txt	Txt
	sh_5454_JD_032219_1_.raw	Raw
	sh_5454_JD_032219_2.raw	Raw

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