Small-molecule-induced DNA damage identifies alternative DNA structures in human genes.
Guanine-rich DNA sequences that can adopt non-Watson-Crick structures in vitro are prevalent in the human genome. Whether such structures normally exist in mammalian cells has, however, been the subject of active research for decades. Here we show that the G-quadruplex-interacting drug pyridostatin promotes growth arrest in human cancer cells by inducing replication- and transcription-dependent DNA damage. A chromatin immunoprecipitation sequencing analysis of the DNA damage marker γH2AX provided the genome-wide distribution of pyridostatin-induced sites of damage and revealed that pyridostatin targets gene bodies containing clusters of sequences with a propensity for G-quadruplex formation. As a result, pyridostatin modulated the expression of these genes, including the proto-oncogene SRC. We observed that pyridostatin reduced SRC protein abundance and SRC-dependent cellular motility in human breast cancer cells, validating SRC as a target of this drug. Our unbiased approach to define genomic sites of action for a drug establishes a framework for discovering functional DNA-drug interactions.
Department of Chemistry, University of Cambridge, Cambridge, UK.
Raphaël
R
Rodriguez
Kyle M
KM
Miller
Josep V
JV
Forment
Charles R
CR
Bradshaw
Mehran
M
Nikan
Sébastien
S
Britton
Tobias
T
Oelschlaegel
Blerta
B
Xhemalce
Shankar
S
Balasubramanian
Stephen P
SP
Jackson
D000634:Aminoquinolines / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D000970:Antineoplastic Agents / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D001943:Breast Neoplasms / Q000235:genetics / Q000473:pathology; D002453:Cell Cycle / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D004247:DNA / Q000737:chemistry* / Q000187:drug effects* / Q000235:genetics; D004249:DNA Damage; D004354:Drug Screening Assays, Antitumor; D054856:G-Quadruplexes / Q000187:drug effects; D006801:Humans; D008970:Molecular Weight; D010848:Picolinic Acids / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D000090063:Proto-Oncogene Mas; D013329:Structure-Activity Relationship; D014407:Tumor Cells, Cultured
D000634:Aminoquinolines; D000970:Antineoplastic Agents; C000715470:MAS1 protein, human; D010848:Picolinic Acids; D000090063:Proto-Oncogene Mas; C567962:pyridostatin; D004247:DNA