The Elg1 Replication Factor C-like Complex Functions in PCNA Unloading during DNA Replication

The ring-shaped complex PCNA coordinates DNA replication, encircling DNA to act as a polymerase clamp and a sliding platform to recruit other replication proteins. PCNA is loaded onto DNA by replication factor C, but it has been unknown how PCNA is removed from DNA when Okazaki fragments are completed or the replication fork terminates. Here we show that the Elg1 replication factor C-like complex (Elg1-RLC) functions in PCNA unloading. Using an improved degron system we show that without Elg1, PCNA accumulates on Saccharomyces cerevisiae chromatin during replication. The accumulated PCNA can be removed from chromatin in vivo by switching on Elg1 expression. We find moreover that treating chromatin with purified Elg1-RLC causes PCNA unloading in vitro. Our results demonstrate that Elg1-RLC functions in unloading of both unmodified and SUMOylated PCNA during DNA replication, while the genome instability of an elg1Δ mutant suggests timely PCNA unloading is critical for chromosome maintenance. [Display omitted] ► PCNA accumulates on chromatin in cells lacking Elg1 ► Expressing Elg1 causes PCNA removal from chromatin in vivo ► Treating chromatin with purified Elg1 complex causes PCNA unloading in vitro ► Elg1 complex functions in unloading of both unmodified and SUMOylated PCNA