Physical and functional interplay between PCNA DNA clamp and Mre11–Rad50 complex from the archaeon Pyrococcus furiosus
|Author(s)||Hogrel Gaelle1, 2, 3, Lu Yang1, 2, 3, Laurent Sebastien1, 2, 3, Henry Etienne1, 2, 3, Etienne Clarisse4, Duy Khanh Phung 4, Dulermo Remi1, 2, 3, Bosse Audrey1, 2, 3, Pluchon Pierre Francois1, 2, 3, Clouet-D'Orval Beatrice4, Flament Didier1, 2, 3|
|Affiliation(s)||1 : IFREMER, UMR6197, Lab Microbiol Environm Extremes, F-29280 Plouzane, France.
2 : Univ Bretagne Occidentale, UMR6197, Lab Microbiol Environm Extremes, F-29280 Plouzane, France.
3 : CNRS, UMR6197, Lab Microbiol Environm Extremes, F-29280 Plouzane, France.
4 : Univ Toulouse, 118 Route Narbonne, F-31062 Toulouse, France.
|Source||Nucleic Acids Research (0305-1048) (Oxford Univ Press), 2018-06 , Vol. 46 , N. 11 , P. 5651-5663|
Several archaeal species prevalent in extreme environments are particularly exposed to factors likely to cause DNA damages. These include hyperthermophilic archaea (HA), living at temperatures >70°C, which arguably have efficient strategies and robust genome guardians to repair DNA damage threatening their genome integrity. In contrast to Eukarya and other archaea, homologous recombination appears to be a vital pathway in HA, and the Mre11–Rad50 complex exerts a broad influence on the initiation of this DNA damage response process. In a previous study, we identified a physical association between the Proliferating Cell Nuclear Antigen (PCNA) and the Mre11–Rad50 (MR) complex. Here, by performing co-immunoprecipitation and SPR analyses, we identified a short motif in the C- terminal portion of Pyrococcus furiosus Mre11 involved in the interaction with PCNA. Through this work, we revealed a PCNA-interaction motif corresponding to a variation on the PIP motif theme which is conserved among Mre11 sequences of Thermococcale species. Additionally, we demonstrated functional interplay in vitro between P. furiosus PCNA and MR enzymatic functions in the DNA end resection process. At physiological ionic strength, PCNA stimulates MR nuclease activities for DNA end resection and promotes an endonucleolytic incision proximal to the 5′ strand of double strand DNA break.