title: AMP-activated protein kinase is involved in the activation of the Fanconi anemia/BRCA pathway in response to DNA interstrand crosslinks.
abstract: Fanconi anemia complementation group (FANC) proteins constitute the Fanconi Anemia (FA)/BRCA pathway that is activated in response to DNA interstrand crosslinks (ICLs). We previously performed yeast two-hybrid screening to identify novel FANC-interacting proteins and discovered that the alpha subunit of AMP-activated protein kinase (AMPKα1) was a candidate binding partner of the FANCG protein, which is a component of the FA nuclear core complex. We confirmed the interaction between AMPKα and both FANCG using co-immunoprecipitation experiments. Additionally, we showed that AMPKα interacted with FANCA, another component of the FA nuclear core complex. AMPKα knockdown in U2OS cells decreased FANCD2 monoubiquitination and nuclear foci formation upon mitomycin C-induced ICLs. Furthermore, AMPKα knockdown enhanced cellular sensitivity to MMC. MMC treatment resulted in an increase in AMPKα phosphorylation/activation, indicating AMPK is involved in the cellular response to ICLs. FANCA was phosphorylated by AMPK at S347 and phosphorylation increased with MMC treatment. MMC-induced FANCD2 monoubiquitination and nuclear foci formation were compromised in a U2OS cell line that stably overexpressed the S347A mutant form of FANCA compared to wild-type FANCA-overexpressing cells, indicating a requirement for FANCA phosphorylation at S347 for proper activation of the FA/BRCA pathway. Our data suggest AMPK is involved in the activation of the FA/BRCA pathway.
authors.affiliation: Cancer Cell and Molecular Biology Branch, Research Institute, National Cancer Center, Ilsandong-gu, Goyang, Gyeonggi, 10408, Korea.
authors.forename: Min Jeong
authors.initials: MJ
authors.lastname: Chun
authors.affiliation: Precision Medicine Branch, Research Institute, National Cancer Center, Ilsandong-gu, Goyang, Gyeonggi, 10408, Korea.
authors.forename: Sunshin
authors.initials: S
authors.lastname: Kim
authors.affiliation: Cancer Cell and Molecular Biology Branch, Research Institute, National Cancer Center, Ilsandong-gu, Goyang, Gyeonggi, 10408, Korea.
authors.forename: Soo Kyung
authors.initials: SK
authors.lastname: Hwang
authors.affiliation: Cancer Cell and Molecular Biology Branch, Research Institute, National Cancer Center, Ilsandong-gu, Goyang, Gyeonggi, 10408, Korea.
authors.forename: Bong Sub
authors.initials: BS
authors.lastname: Kim
authors.affiliation: Cancer Cell and Molecular Biology Branch, Research Institute, National Cancer Center, Ilsandong-gu, Goyang, Gyeonggi, 10408, Korea.
authors.forename: Hyoun Geun
authors.initials: HG
authors.lastname: Kim
authors.affiliation: Cancer Cell and Molecular Biology Branch, Research Institute, National Cancer Center, Ilsandong-gu, Goyang, Gyeonggi, 10408, Korea.
authors.forename: Hae In
authors.initials: HI
authors.lastname: Choi
authors.affiliation: Cancer Cell and Molecular Biology Branch, Research Institute, National Cancer Center, Ilsandong-gu, Goyang, Gyeonggi, 10408, Korea.
authors.forename: Jong Heon
authors.initials: JH
authors.lastname: Kim
authors.affiliation: Precision Medicine Branch, Research Institute, National Cancer Center, Ilsandong-gu, Goyang, Gyeonggi, 10408, Korea.
authors.forename: Sung Ho
authors.initials: SH
authors.lastname: Goh
authors.affiliation: Cancer Cell and Molecular Biology Branch, Research Institute, National Cancer Center, Ilsandong-gu, Goyang, Gyeonggi, 10408, Korea.
authors.forename: Chang-Hun
authors.initials: CH
authors.lastname: Lee
mesh_terms: D055372:AMP-Activated Protein Kinases / Q000378:metabolism*; D045744:Cell Line, Tumor; D004249:DNA Damage; D052217:Fanconi Anemia Complementation Group A Protein / Q000378:metabolism*; D052241:Fanconi Anemia Complementation Group G Protein / Q000378:metabolism*; D006801:Humans
chemical_list: C498471:FANCA protein, human; C499909:FANCG protein, human; D052217:Fanconi Anemia Complementation Group A Protein; D052241:Fanconi Anemia Complementation Group G Protein; D055372:AMP-Activated Protein Kinases
keywords: AMP-activated protein kinase; DNA interstrand crosslinks; FANCA; Fanconi anemia (FA)/BRCA pathway; phosphorylation