WWP2 ubiquitylates RNA polymerase II for DNA-PK-dependent transcription arrest and repair at DNA breaks

• Published in: Genes & development Vol. 33; no. 11-12; pp. 684 - 704
• Main Authors: Caron, Pierre, Pankotai, Tibor, Wiegant, Wouter W, Tollenaere, Maxim A X, Furst, Audrey, Bonhomme, Celine, Helfricht, Angela, de Groot, Anton, Pastink, Albert, Vertegaal, Alfred C O, Luijsterburg, Martijn S, Soutoglou, Evi, van Attikum, Haico
• Format: Journal Article
• Language: English
• Published: United States Cold Spring Harbor Laboratory Press 01-06-2019
• Subjects: Cell Line, Transformed; Cell Line, Tumor; DNA Breaks, Double-Stranded; DNA End-Joining Repair; DNA-Activated Protein Kinase - metabolism; DNA-Directed RNA Polymerases - metabolism; Humans; Life Sciences; Nuclear Proteins - metabolism; Proteasome Endopeptidase Complex - metabolism; Research Paper; RNA Polymerase II - metabolism; Transcription, Genetic; Ubiquitin-Protein Ligases - metabolism; Ubiquitination; transcription silencing; DNA double-strand break repair; DNA-PK; RNAPII ubiquitylation; WWP2 HECT E3 ubiquitin ligase
• ISSN: 0890-9369; 1549-5477
• DOI: 10.1101/gad.321943.118

DNA double-strand breaks (DSBs) at RNA polymerase II (RNAPII) transcribed genes lead to inhibition of transcription. The DNA-dependent protein kinase (DNA-PK) complex plays a pivotal role in transcription inhibition at DSBs by stimulating proteasome-dependent eviction of RNAPII at these lesions. How DNA-PK triggers RNAPII eviction to inhibit transcription at DSBs remains unclear. Here we show that the HECT E3 ubiquitin ligase WWP2 associates with components of the DNA-PK and RNAPII complexes and is recruited to DSBs at RNAPII transcribed genes. In response to DSBs, WWP2 targets the RNAPII subunit RPB1 for K48-linked ubiquitylation, thereby driving DNA-PK- and proteasome-dependent eviction of RNAPII. The lack of WWP2 or expression of nonubiquitylatable RPB1 abrogates the binding of nonhomologous end joining (NHEJ) factors, including DNA-PK and XRCC4/DNA ligase IV, and impairs DSB repair. These findings suggest that WWP2 operates in a DNA-PK-dependent shutoff circuitry for RNAPII clearance that promotes DSB repair by protecting the NHEJ machinery from collision with the transcription machinery.