Association with Aurora-A Controls N-MYC-Dependent Promoter Escape and Pause Release of RNA Polymerase II during the Cell Cycle

• Published in: Cell reports (Cambridge) 2017-12, Vol.21 (12), p.3483-3497
• Main Authors: Büchel, Gabriele, Carstensen, Anne, Mak, Ka-Yan, Roeschert, Isabelle, Leen, Eoin, Sumara, Olga, Hofstetter, Julia, Herold, Steffi, Kalb, Jacqueline, Baluapuri, Apoorva, Poon, Evon, Kwok, Colin, Chesler, Louis, Maric, Hans Michael, Rickman, David S., Wolf, Elmar, Bayliss, Richard, Walz, Susanne, Eilers, Martin
• Format: Article
• Language: English
• Subjects: Aurora Kinase A - metabolism; Aurora-A; Cell Cycle Proteins; Cell Line, Tumor; DNA Topoisomerases, Type II - metabolism; DNA, Intergenic - metabolism; DNA-Binding Proteins; Humans; MYC; N-MYC; N-Myc Proto-Oncogene Protein - metabolism; neuroblastoma; Nuclear Proteins - metabolism; pause release; Phosphoproteins - metabolism; Promoter Regions, Genetic; Protein Binding; RAD21; RNA Polymerase II - genetics; RNA Polymerase II - metabolism; S Phase; TFIIIC; Transcription Elongation, Genetic; Transcription Factors, TFIII - metabolism
• ISSN: 2211-1247; 2211-1247
• DOI: 10.1016/j.celrep.2017.11.090

MYC proteins bind globally to active promoters and promote transcriptional elongation by RNA polymerase II (Pol II). To identify effector proteins that mediate this function, we performed mass spectrometry on N-MYC complexes in neuroblastoma cells. The analysis shows that N-MYC forms complexes with TFIIIC, TOP2A, and RAD21, a subunit of cohesin. N-MYC and TFIIIC bind to overlapping sites in thousands of Pol II promoters and intergenic regions. TFIIIC promotes association of RAD21 with N-MYC target sites and is required for N-MYC-dependent promoter escape and pause release of Pol II. Aurora-A competes with binding of TFIIIC and RAD21 to N-MYC in vitro and antagonizes association of TOP2A, TFIIIC, and RAD21 with N-MYC during S phase, blocking N-MYC-dependent release of Pol II from the promoter. Inhibition of Aurora-A in S phase restores RAD21 and TFIIIC binding to chromatin and partially restores N-MYC-dependent transcriptional elongation. We propose that complex formation with Aurora-A controls N-MYC function during the cell cycle. [Display omitted] •N-MYC forms complexes with TFIIIC, RAD21, and TOP2A•TFIIIC recruits RAD21 and is required for N-MYC-dependent pause release of Pol II•Aurora-A displaces TFIIIC, TOP2A, and RAD21 from N-MYC during S phase•Aurora-A inhibits pause release of Pol II during S phase Büchel et al. demonstrate that N-MYC forms complexes with TFIIIC, TOP2A, and RAD21. Aurora-A competes with TFIIIC and RAD21 for binding to N-MYC, and Aurora-A displaces the three proteins from N-MYC during S phase. As consequence, N-MYC-dependent pause release is inhibited during S phase, preventing activation of the ATR checkpoint kinase.