Regulation of PCNA cycling on replicating DNA by RFC and RFC-like complexes.
Replication-Factor-C (RFC) and RFC-like complexes (RLCs) mediate chromatin engagement of the proliferating cell nuclear antigen (PCNA). It remains controversial how RFC and RLCs cooperate to regulate PCNA loading and unloading. Here, we show the distinct PCNA loading or unloading activity of each clamp loader. ATAD5-RLC possesses the potent PCNA unloading ... activity. ATPase motif and collar domain of ATAD5 are crucial for the unloading activity. DNA structures did not affect PCNA unloading activity of ATAD5-RLC. ATAD5-RLC could unload ubiquitinated PCNA. Through single molecule measurements, we reveal that ATAD5-RLC unloaded PCNA through one intermediate state before ATP hydrolysis. RFC loaded PCNA through two intermediate states on DNA, separated by ATP hydrolysis. Replication proteins such as Fen1 could inhibit the PCNA unloading activity of Elg1-RLC, a yeast homolog of ATAD5-RLC in vitro. Our findings provide molecular insights into how PCNA is released from chromatin to finalize DNA replication/repair.
Mesh Terms:
ATPases Associated with Diverse Cellular Activities, Adenosine Triphosphatases, Adenosine Triphosphate, Carrier Proteins, Chromatin, DNA, DNA Replication, DNA-Binding Proteins, Flap Endonucleases, Humans, Hydrolysis, Proliferating Cell Nuclear Antigen, Replication Protein C, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins
ATPases Associated with Diverse Cellular Activities, Adenosine Triphosphatases, Adenosine Triphosphate, Carrier Proteins, Chromatin, DNA, DNA Replication, DNA-Binding Proteins, Flap Endonucleases, Humans, Hydrolysis, Proliferating Cell Nuclear Antigen, Replication Protein C, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins
Nat Commun
Date: Dec. 03, 2018
PubMed ID: 31160570
View in: Pubmed Google Scholar
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