Requirements for PCNA monoubiquitination in human cell-free extracts.
The Rad6/Rad18-dependent monoubiquitination of PCNA plays a crucial role in regulating replication past DNA damage in eukaryotic cells. We show here that in human cell-free extracts, efficient PCNA monoubiquitination requires both the synthesis of relatively long DNA tracts and polymerase idling or stalling at sites of DNA modification or DNA secondary structures. This dual dependency suggests a dynamic process in which, following initiation, the DNA synthesizing complex undergoes modifications that make it competent as a mediator for the activation of the Rad6/Rad18 pathway.
Institut Gilbert Laustriat, Biomolécules, Biotechnologie, Innovations thérapeutiques,UMR 7175 - LC1 - CNRS/ULP, ESBS, Bld Sébastien Brandt, BP 10413, Illkirch 67412, France.
Valérie
V
Schmutz
Jérôme
J
Wagner
Régine
R
Janel-Bintz
Robert P P
RP
Fuchs
Agnès M
AM
Cordonnier
D002474:Cell-Free System; D004261:DNA Replication; D006367:HeLa Cells; D006801:Humans; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism*; D025801:Ubiquitin / Q000378:metabolism*
D018809:Proliferating Cell Nuclear Antigen; D025801:Ubiquitin