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Su Hyung, Park
유전체 항상성 연구단
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Characterization of subcellular localization of eukaryotic clamp loader/unloader and its regulatory mechanism

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Title
Characterization of subcellular localization of eukaryotic clamp loader/unloader and its regulatory mechanism
Author(s)
Su Hyung Park; Seong-jung Kim; Kyungjae Myung; Kyoo-young Lee
Publication Date
2021-11
Journal
Scientific Reports, v.11, no.1
Publisher
Nature Research
Abstract
© 2021, The Author(s).Proliferating cell nuclear antigen (PCNA) plays a critical role as a processivity clamp for eukaryotic DNA polymerases and a binding platform for many DNA replication and repair proteins. The enzymatic activities of PCNA loading and unloading have been studied extensively in vitro. However, the subcellular locations of PCNA loaders, replication complex C (RFC) and CTF18-RFC-like-complex (RLC), and PCNA unloader ATAD5-RLC remain elusive, and the role of their subunits RFC2-5 is unknown. Here we used protein fractionation to determine the subcellular localization of RFC and RLCs and affinity purification to find molecular requirements for the newly defined location. All RFC/RLC proteins were detected in the nuclease-resistant pellet fraction. RFC1 and ATAD5 were not detected in the non-ionic detergent-soluble and nuclease-susceptible chromatin fractions, independent of cell cycle or exogenous DNA damage. We found that small RFC proteins contribute to maintaining protein levels of the RFC/RLCs. RFC1, ATAD5, and RFC4 co-immunoprecipitated with lamina-associated polypeptide 2 (LAP2) α which regulates intranuclear lamin A/C. LAP2α knockout consistently reduced detection of RFC/RLCs in the pellet fraction, while marginally affecting total protein levels. Our findings strongly suggest that PCNA-mediated DNA transaction occurs through regulatory machinery associated with nuclear structures, such as the nuclear matrix.
URI
https://pr.ibs.re.kr/handle/8788114/10916
DOI
10.1038/s41598-021-01336-w
ISSN
2045-2322
Appears in Collections:
Center for Genomic Integrity(유전체 항상성 연구단) > 1. Journal Papers (저널논문)
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