Dataset Information

Analysis of Rev1-dependent replication dynamics using isolation of proteins on nascent DNA

ABSTRACT: To study the influence of the Rev1 translesion synthesis enzyme on fork dynamics, we performed isolation of proteins on nascent DNA coupled with mass spectrometry (iPOND-MS). Wild-type and REV1-KO HEK293 Flp-In™/T-REx™ cells were used. Treatment with the G-quadruplex stabilizing agent pyridostatin (PDS; 10 micromolar for either 15 or 60 minutes) was used to examine the Rev1-dependent response to G4-induced replication stress. A thymidine pulse experiment was used to ensure that replication proteins were enriched in the samples pulsed with 5-ethynyl-2ʹ-deoxyuridine (EdU). The resulting samples were analyzed using liquid chromatography mass spectrometry (LC-MS). Label-free quantitative analysis of peptide abundance was used to determine changes in response to Rev1 deletion and treatment with PDS.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Dennis Province

LAB HEAD: Robert L. Eoff

PROVIDER: PXD061281 | Pride | 2026-06-15

REPOSITORIES: Pride

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Publications

Human REV1 interacts with DHX36 to promote replication and tolerance of G-quadruplex DNA.

Ketkar Amit A Paxton Bethany C BC Zuniga Oscar E OE Sewilam Reham S RS Morales Martin M Schaller John J McCollum Rowan J RJ Jackson Kathleen E KE Lowran Kaitlin K Paul Alyssa A Patel Meera M Seenivasan Sreevatsav S McCrury Mason M Al-Anbaky Qudes Q Maddukuri Leena L Kendrick Samantha S Wu Colin G CG Gunderson Julie E C JEC Eoff Robert L RL

Nucleic acids research 20260601 11

G-quadruplex DNA is a barrier to replication, but how the replisome couples G4 bypass with fork progression is not well-defined. Here, we establish that REV1 is central to coordination of G4 resolution, replication fidelity, and tolerance of G4 stabilization. REV1 loss switched fork elongation to a PrimPol-driven mechanism and resulted in defective ssDNA gap suppression in cells treated with pyridostatin (PDS). Mutagenic G4 replication on the leading strand was more impacted by REV1 loss than la ...[more]

PMID: 42258547

			Action	DRS
	Eoff_100821_GPF6_1.mgf	Mgf
	Eoff_100821_GPF6_1.mzML	Mzml
	Eoff_100821_GPF6_2.mgf	Mgf
	Eoff_100821_GPF6_2.mzML	Mzml
	Eoff_100821_GPF6_3.mgf	Mgf

Dataset Information

Analysis of Rev1-dependent replication dynamics using isolation of proteins on nascent DNA

Dataset's files

Publications

Human REV1 interacts with DHX36 to promote replication and tolerance of G-quadruplex DNA.

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