Journal Article

High speed of fork progression induces DNA replication stress and genomic instability

Lee, MyungHee, Strauss, Robert, Maya-Mendoza, Apolinar, Bartek, Jiri, Moudry, Pavel, Merchut-Maya, Joanna Maria

2018

Overview

Accurate replication of DNA requires stringent regulation to ensure genome integrity. In human cells, thousands of origins of replication are coordinately activated during S phase, and the velocity of replication forks is adjusted to fully replicate DNA in pace with the cell cycle 1 . Replication stress induces fork stalling and fuels genome instability 2 . The mechanistic basis of replication stress remains poorly understood despite its emerging role in promoting cancer 2 . Here we show that inhibition of poly(ADP-ribose) polymerase (PARP) increases the speed of fork elongation and does not cause fork stalling, which is in contrast to the accepted model in which inhibitors of PARP induce fork stalling and collapse 3 . Aberrant acceleration of fork progression by 40% above the normal velocity leads to DNA damage. Depletion of the treslin or MTBP proteins, which are involved in origin firing, also increases fork speed above the tolerated threshold, and induces the DNA damage response pathway. Mechanistically, we show that poly(ADP-ribosyl)ation (PARylation) and the PCNA interactor p21 Cip1 (p21) are crucial modulators of fork progression. PARylation and p21 act as suppressors of fork speed in a coordinated regulatory network that is orchestrated by the PARP1 and p53 proteins. Moreover, at the fork level, PARylation acts as a sensor of replication stress. During PARP inhibition, DNA lesions that induce fork arrest and are normally resolved or repaired remain unrecognized by the replication machinery. Conceptually, our results show that accelerated replication fork progression represents a general mechanism that triggers replication stress and the DNA damage response. Our findings contribute to a better understanding of the mechanism of fork speed control, with implications for genomic (in)stability and rational cancer treatment. Inhibition of PARP is shown to accelerate the speed of replication fork elongation, which prevents fork stalling and induces DNA damage, with implications for genomic instability and cancer treatment.

• Publisher: Nature Publishing Group UK,Nature Publishing Group
• Subject: 13/31 / 14/19 / 631/337/151/2356 / 631/67/1059/99 / 82/29 / Adenosine diphosphate / ADP-ribosylation / Analysis / Cancer / Cancer research / Cell cycle / Cell Line, Tumor / Cells (Biology) / Chromosome Structures - drug effects / Control stability / Cyclin-dependent kinase inhibitor p21 / Cyclin-Dependent Kinase Inhibitor p21 - metabolism / Damage tolerance / Deoxyribonucleic acid / DNA / DNA biosynthesis / DNA damage / DNA Damage - drug effects / DNA repair / DNA replication / DNA Replication - drug effects / DNA Replication - physiology / Elongation / Gene expression / Genetic research / Genomes / Genomic instability / Genomic Instability - drug effects / Genomics / Humanities and Social Sciences / Humans / Instability / Kinases / Lesions / Letter / Modulators / Monosaccharides / multidisciplinary / p53 Protein / Phosphorylation / Phthalazines - pharmacology / Piperazines - pharmacology / Poly (ADP-Ribose) Polymerase-1 - antagonists & inhibitors / Poly (ADP-Ribose) Polymerase-1 - metabolism / Poly(ADP-ribose) / Poly(ADP-ribose) polymerase / Poly(ADP-ribose) Polymerase Inhibitors - pharmacology / Proliferating cell nuclear antigen / Proteins / Replication / Replication forks / Replication origins / Ribose / S phase / Science / Science (multidisciplinary) / Speed control / Stability / Stalling / Stresses / Suppressors / Time Factors / Tumor proteins / Tumor Suppressor Protein p53 - metabolism / Velocity