AKAP12 mediates PKA-induced phosphorylation of ATR to enhance nucleotide excision repair.
Loss-of-function in melanocortin 1 receptor (MC1R), a GSprotein-coupled receptor that regulates signal transduction through cAMP and protein kinase A (PKA) in melanocytes, is a major inherited melanoma risk factor. Herein, we report a novel cAMP-mediated response for sensing and responding to UV-induced DNA damage regulated by A-kinase-anchoring protein 12 (AKAP12). AKAP12 is identified as a necessary participant in PKA-mediated phosphorylation of ataxia telangiectasia mutated and Rad3-related (ATR) at S435, a post-translational event required for cAMP-enhanced nucleotide excision repair (NER). Moreover, UV exposure promotes ATR-directed phosphorylation of AKAP12 at S732, which promotes nuclear translocation of AKAP12-ATR-pS435. This complex subsequently recruits XPA to UV DNA damage and enhances 5' strand incision. Preventing AKAP12's interaction with PKA or with ATR abrogates ATR-pS435 accumulation, delays recruitment of XPA to UV-damaged DNA, impairs NER and increases UV-induced mutagenesis. Our results define a critical role for AKAP12 as an UV-inducible scaffold for PKA-mediated ATR phosphorylation, and identify a repair complex consisting of AKAP12-ATR-pS435-XPA at photodamage, which is essential for cAMP-enhanced NER.
Markey Cancer Center, University of Kentucky College of Medicine, Lexington, KY 40536, USA.
Stuart G
SG
Jarrett
Markey Cancer Center, University of Kentucky College of Medicine, Lexington, KY 40536, USA.|Department of Physiology, University of Kentucky College of Medicine, Lexington, KY 40536, USA.
Erin M
EM
Wolf Horrell
Markey Cancer Center, University of Kentucky College of Medicine, Lexington, KY 40536, USA jdorazio@uky.edu.|Department of Physiology, University of Kentucky College of Medicine, Lexington, KY 40536, USA.|Department of Toxicology and Cancer Biology, University of Kentucky College of Medicine, Lexington, KY 40536, USA.|Department of Pharmacology and Nutritional Science, University of Kentucky College of Medicine, Lexington, KY 40536, USA.|Department of Pediatrics, University of Kentucky College of Medicine, Lexington, KY 40536, USA.
John A
JA
D'Orazio
D054758:A Kinase Anchor Proteins / Q000502:physiology*; D064007:Ataxia Telangiectasia Mutated Proteins / Q000378:metabolism; D018797:Cell Cycle Proteins / Q000502:physiology*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000502:physiology*; D004249:DNA Damage; D004260:DNA Repair; D057809:HEK293 Cells; D006801:Humans; D007700:Kinetics; D016296:Mutagenesis; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D021381:Protein Transport; D051760:Xeroderma Pigmentosum Group A Protein / Q000378:metabolism
D054758:A Kinase Anchor Proteins; C491162:AKAP12 protein, human; D018797:Cell Cycle Proteins; C497314:XPA protein, human; D051760:Xeroderma Pigmentosum Group A Protein; C576597:ATR protein, human; D064007:Ataxia Telangiectasia Mutated Proteins; D017868:Cyclic AMP-Dependent Protein Kinases
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