title: A neomorphic cancer cell-specific role of MAGE-A4 in trans-lesion synthesis.
abstract: Trans-lesion synthesis (TLS) is an important DNA-damage tolerance mechanism that permits ongoing DNA synthesis in cells harbouring damaged genomes. The E3 ubiquitin ligase RAD18 activates TLS by promoting recruitment of Y-family DNA polymerases to sites of DNA-damage-induced replication fork stalling. Here we identify the cancer/testes antigen melanoma antigen-A4 (MAGE-A4) as a tumour cell-specific RAD18-binding partner and an activator of TLS. MAGE-A4 depletion from MAGE-A4-expressing cancer cells destabilizes RAD18. Conversely, ectopic expression of MAGE-A4 (in cell lines lacking endogenous MAGE-A4) promotes RAD18 stability. DNA-damage-induced mono-ubiquitination of the RAD18 substrate PCNA is attenuated by MAGE-A4 silencing. MAGE-A4-depleted cells fail to resume DNA synthesis normally following ultraviolet irradiation and accumulate γH2AX, thereby recapitulating major hallmarks of TLS deficiency. Taken together, these results demonstrate a mechanism by which reprogramming of ubiquitin signalling in cancer cells can influence DNA damage tolerance and probably contribute to an altered genomic landscape.
authors.affiliation: Department of Pathology and Laboratory Medicine, University of North Carolina at Chapel Hill, 101 Manning Drive, 614 Brinkhous-Bullitt Building, Chapel Hill, North Carolina 27599, USA.
authors.forename: Yanzhe
authors.initials: Y
authors.lastname: Gao
authors.affiliation: Department of Pathology and Laboratory Medicine, University of North Carolina at Chapel Hill, 101 Manning Drive, 614 Brinkhous-Bullitt Building, Chapel Hill, North Carolina 27599, USA.|Curriculum in Toxicology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, USA.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Mutter-Rottmayer
authors.affiliation: Department of Pathology and Laboratory Medicine, University of North Carolina at Chapel Hill, 101 Manning Drive, 614 Brinkhous-Bullitt Building, Chapel Hill, North Carolina 27599, USA.|Curriculum in Genetics and Molecular Biology, University of North Carolina, Chapel Hill, North Carolina 27599, USA.
authors.forename: Alicia M
authors.initials: AM
authors.lastname: Greenwalt
authors.affiliation: Department of Computer Science, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, USA.
authors.forename: Dennis
authors.initials: D
authors.lastname: Goldfarb
authors.affiliation: Department of Cell Biology and Physiology, Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, USA.
authors.forename: Feng
authors.initials: F
authors.lastname: Yan
authors.affiliation: Department of Pathology and Laboratory Medicine, University of North Carolina at Chapel Hill, 101 Manning Drive, 614 Brinkhous-Bullitt Building, Chapel Hill, North Carolina 27599, USA.
authors.forename: Yang
authors.initials: Y
authors.lastname: Yang
authors.affiliation: Department of Pathology and Laboratory Medicine, University of North Carolina at Chapel Hill, 101 Manning Drive, 614 Brinkhous-Bullitt Building, Chapel Hill, North Carolina 27599, USA.|Department of Pharmacology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, USA.
authors.forename: Raquel C
authors.initials: RC
authors.lastname: Martinez-Chacin
authors.affiliation: Center For Integrative Chemical Biology and Drug Discovery, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, USA.
authors.forename: Kenneth H
authors.initials: KH
authors.lastname: Pearce
authors.affiliation: Division of Cell Maintenance, Institute of Molecular Embryology and Genetics (IMEG), Kumamoto University, Honjo 2-2-1, Kumamoto 860-0811, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Tateishi
authors.affiliation: Department of Computer Science, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, USA.|Department of Cell Biology and Physiology, Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, USA.
authors.forename: Michael B
authors.initials: MB
authors.lastname: Major
authors.affiliation: Department of Pathology and Laboratory Medicine, University of North Carolina at Chapel Hill, 101 Manning Drive, 614 Brinkhous-Bullitt Building, Chapel Hill, North Carolina 27599, USA.|Curriculum in Toxicology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, USA.|Curriculum in Genetics and Molecular Biology, University of North Carolina, Chapel Hill, North Carolina 27599, USA.
authors.forename: Cyrus
authors.initials: C
authors.lastname: Vaziri
mesh_terms: D000072283:A549 Cells; D000818:Animals; D000951:Antigens, Neoplasm / Q000235:genetics* / Q000378:metabolism; D002460:Cell Line; D045744:Cell Line, Tumor; D003001:Cloning, Molecular; D004249:DNA Damage; D004260:DNA Repair; D004261:DNA Replication; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism / Q000528:radiation effects; D015972:Gene Expression Regulation, Neoplastic; D005822:Genetic Vectors / Q000737:chemistry / Q000378:metabolism; D045325:HCT116 Cells; D006367:HeLa Cells; D006657:Histones / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D051379:Mice; D009363:Neoplasm Proteins / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D044763:Ubiquitin-Conjugating Enzymes / Q000235:genetics / Q000378:metabolism; D044767:Ubiquitin-Protein Ligases / Q000235:genetics* / Q000378:metabolism; D014466:Ultraviolet Rays
chemical_list: D000951:Antigens, Neoplasm; D004268:DNA-Binding Proteins; C468783:H2AX protein, human; D006657:Histones; C095204:MAGEA4 protein, human; D009363:Neoplasm Proteins; C485954:RAD18 protein, human; D011994:Recombinant Proteins; C471545:UBE2A protein, human; D044763:Ubiquitin-Conjugating Enzymes; D044767:Ubiquitin-Protein Ligases