Proteomics

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AP-MS of replisomes from S. cerevisiae in S phase and after unscheduled G1 replication

ABSTRACT: Interfering with once-per-cell-cycle regulation of DNA replication initiation generates genome instability through over-replication and has been linked to early stages of cancer development. Here, we engineered different systems in budding yeast to induce over-replication in G1 and investigated its characteristics and consequences within the same cell cycle. To study replisome composition, we immunopurified replisomes via a GFP-tag on the Psf2 subunit of the GINS complex from S phase in the presence and absence of hydroxyurea and after unscheduled replication initiation in G1.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Saccharomyces Cerevisiae (baker's Yeast)

SUBMITTER: Barbara Steigenberger  

LAB HEAD: Boris Pfander

PROVIDER: PXD035629 | Pride | 2023-03-11

REPOSITORIES: Pride

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Publications

Unscheduled DNA replication in G1 causes genome instability and damage signatures indicative of replication collisions.

Reusswig Karl-Uwe KU   Bittmann Julia J   Peritore Martina M   Courtes Mathilde M   Pardo Benjamin B   Wierer Michael M   Mann Matthias M   Pfander Boris B  

Nature communications 20221118 1

DNA replicates once per cell cycle. Interfering with the regulation of DNA replication initiation generates genome instability through over-replication and has been linked to early stages of cancer development. Here, we engineer genetic systems in budding yeast to induce unscheduled replication in a G1-like cell cycle state. Unscheduled G1 replication initiates at canonical S-phase origins. We quantifiy the composition of replisomes in G1- and S-phase and identified firing factors, polymerase α,  ...[more]

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