Abstract

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.

Replication-Factor-C (RFC) and RFC-like complexes (RLCs) mediate chromatin engagement of the proliferating cell nuclear antigen (PCNA). Here authors use biochemical and single molecule measurements to show that ATAD5-RLC has the most potent PCNA unloading activity and forms structurally distinct intermediates compared to RFC-PCNA.

Details

Title
Regulation of PCNA cycling on replicating DNA by RFC and RFC-like complexes
Author
Mi-Sun, Kang 1   VIAFID ORCID Logo  ; Ryu Eunjin 2 ; Seung-Won, Lee 3 ; Park, Jieun 1 ; Ha Na Young 1 ; Ra Jae Sun 1 ; Kim, Yeong Jae 2 ; Kim, Jinwoo 2 ; Abdel-Rahman, Mohamed 4 ; Park, Su Hyung 1 ; Kyoo-young, Lee 1 ; Kim Hajin 2 ; Kang Sukhyun 1 ; Myung Kyungjae 2 

1  Center for Genomic Integrity, Institute for Basic Science, Ulsan, Republic of Korea (GRID:grid.410720.0) (ISNI:0000 0004 1784 4496) 
2  Center for Genomic Integrity, Institute for Basic Science, Ulsan, Republic of Korea (GRID:grid.410720.0) (ISNI:0000 0004 1784 4496); School of Life Sciences, Ulsan National Institute of Science and Technology, Ulsan, Republic of Korea (GRID:grid.42687.3f) (ISNI:0000 0004 0381 814X) 
3  School of Life Sciences, Ulsan National Institute of Science and Technology, Ulsan, Republic of Korea (GRID:grid.42687.3f) (ISNI:0000 0004 0381 814X) 
4  Ohio State University Comprehensive Cancer Center, Department of Ophthalmology, Columbus, USA (GRID:grid.261331.4) (ISNI:0000 0001 2285 7943) 
Publication year
2019
Publication date
2019
Publisher
Nature Publishing Group
e-ISSN
20411723
Source type
Scholarly Journal
Language of publication
English
DOI
https://doi.org/10.1038/s41467-019-10376-w
ProQuest document ID
2233826599
Copyright
© The Author(s) 2019. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.