Flap Endonuclease 1 Limits Telomere Fragility on the Leading Strand.
The existence of redundant replication and repair systems that ensure genome stability underscores the importance of faithful DNA replication. Nowhere is this complexity more evident than in challenging DNA templates, including highly repetitive or transcribed sequences. Here, we demonstrate that flap endonuclease 1 (FEN1), a canonical lagging strand DNA replication protein, is required for normal, complete leading strand replication at telomeres. We find that the loss of FEN1 nuclease activity, but not DNA repair activities, results in leading strand-specific telomere fragility. Furthermore, we show that FEN1 depletion-induced telomere fragility is increased by RNA polymerase II inhibition and is rescued by ectopic RNase H1 expression. These data suggest that FEN1 limits leading strand-specific telomere fragility by processing RNA:DNA hybrid/flap intermediates that arise from co-directional collisions occurring between the replisome and RNA polymerase. Our data reveal the first molecular mechanism for leading strand-specific telomere fragility and the first known role for FEN1 in leading strand DNA replication. Because FEN1 mutations have been identified in human cancers, our findings raise the possibility that unresolved RNA:DNA hybrid structures contribute to the genomic instability associated with cancer.
From the Departments of Cell Biology and Physiology and Integrating Communications within the Cancer Environment Institute, Washington University School of Medicine, Saint Louis, Missouri 63110.
Daniel C
DC
Teasley
From the Departments of Cell Biology and Physiology and Integrating Communications within the Cancer Environment Institute, Washington University School of Medicine, Saint Louis, Missouri 63110.
Shankar
S
Parajuli
From the Departments of Cell Biology and Physiology and Integrating Communications within the Cancer Environment Institute, Washington University School of Medicine, Saint Louis, Missouri 63110.
Mai
M
Nguyen
From the Departments of Cell Biology and Physiology and Integrating Communications within the Cancer Environment Institute, Washington University School of Medicine, Saint Louis, Missouri 63110.
Hayley R
HR
Moore
From the Departments of Cell Biology and Physiology and Integrating Communications within the Cancer Environment Institute, Washington University School of Medicine, Saint Louis, Missouri 63110.
Elise
E
Alspach
From the Departments of Cell Biology and Physiology and.
Ying Jie
YJ
Lock
From the Departments of Cell Biology and Physiology and.
Yuchi
Y
Honaker
From the Departments of Cell Biology and Physiology and.
Abhishek
A
Saharia
From the Departments of Cell Biology and Physiology and Medicine.
Helen
H
Piwnica-Worms
From the Departments of Cell Biology and Physiology and Integrating Communications within the Cancer Environment Institute, Washington University School of Medicine, Saint Louis, Missouri 63110 Medicine, sheila.stewart@wustl.edu.
Sheila A
SA
Stewart
D015153:Blotting, Western; D004249:DNA Damage; D004261:DNA Replication; D045585:Flap Endonucleases / Q000235:genetics / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016615:Telomere; D014158:Transcription, Genetic
D045585:Flap Endonucleases
DNA replication; FEN1; cancer; genomic instability; telomere; transcription