MDC1 maintains active elongation complexes of RNA polymerase II
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The role of MDC1 in the DNA damage response has been extensively studied; however, its impact on other cellular processes is not well understood. Here, we describe the role of MDC1 in transcription as a regulator of RNA polymerase II (RNAPII). Depletion of MDC1 causes a genome-wide reduction in the abundance of actively engaged RNAPII elongation complexes throughout the gene body of protein-encoding genes under unperturbed conditions. Decreased engaged RNAPII subsequently alters the assembly of the spliceosome complex on chromatin, leading to changes in pre-mRNA splicing. Mechanistically, the S/TQ domain of MDC1 modulates RNAPII-mediated transcription. Upon genotoxic stress, MDC1 promotes the abundance of engaged RNAPII complexes at DNA breaks, thereby stimulating nascent transcription at the damaged sites. Of clinical relevance, cancer cells lacking MDC1 display hypersensitivity to RNAPII inhibitors. Overall, we unveil a role of MDC1 in RNAPII-mediated transcription with potential implications for cancer treatment.
| Originalsprog | Engelsk |
|---|---|
| Artikelnummer | 111979 |
| Tidsskrift | Cell Reports |
| Vol/bind | 42 |
| Udgave nummer | 1 |
| Antal sider | 28 |
| ISSN | 2211-1247 |
| DOI | |
| Status | Udgivet - 2023 |
Bibliografisk note
Funding Information:
We thank Dr. Junjie Chen for providing the HA-tagged MDC1 WT and deletion mutants ( Figure 4 A). We would like to thank Dr. Fabrizio d’Adda di Fagagna and Dr. Matteo Cabrini for the scientific discussions that improved the present study. J.B. and the members of his groups were funded by the European Union’s Horizon 2020 research and innovation program under the Marie Skłodowska-Curie grant agreement 722729 , by grants from the Danish Cancer Society ( R167-A11068 and R204-A12617-B153 ), the Lundbeck Foundation ( R266-2017-4289 ), the Novo Nordisk Foundation ( NNF20OC0060590 ), the Swedish Research Council ( VR-MH 2014-46602-117891-30 ), the Danish Council for Independent Research ( DFF-7016-00313 ), and the Danish National Research Foundation (project CARD, DNRF 125 ). K.W. was supported by Japan Society for the Promotion of Science ( JSPS ) KAKENHI grants JP19K23927 and JP20K07578 . P.G. is funded by a Lundbeck postdoctoral fellowship ( R322-2019-2577 ). A.M.-M. was funded by the Danish Cancer Society ( R302-A17590 ). D.H.L. and Z.G. were supported by the Danish Cancer Society, Danish Cancer Society-Knæk Cancer ( R311-A18224 ), the Independent Research Fund Denmark ( 8045-00057A ), and the Novo Nordisk Foundation ( NNF18OC0052647 ). Work in the lab of L.H.G. is funded by a Hallas-Møller emerging investigator grant from the Novo Nordisk Foundation ( NNF20OC0059959 ) and a Sapere Aude grant from Independent Research Fund Denmark ( 0165-00092B ). Q.T. is funded by a Lundbeck postdoctoral fellowship ( R380-2021-1284 ).
Funding Information:
We thank Dr. Junjie Chen for providing the HA-tagged MDC1 WT and deletion mutants (Figure 4A). We would like to thank Dr. Fabrizio d'Adda di Fagagna and Dr. Matteo Cabrini for the scientific discussions that improved the present study. J.B. and the members of his groups were funded by the European Union's Horizon 2020 research and innovation program under the Marie Skłodowska-Curie grant agreement 722729, by grants from the Danish Cancer Society (R167-A11068 and R204-A12617-B153), the Lundbeck Foundation (R266-2017-4289), the Novo Nordisk Foundation (NNF20OC0060590), the Swedish Research Council (VR-MH 2014-46602-117891-30), the Danish Council for Independent Research (DFF-7016-00313), and the Danish National Research Foundation (project CARD, DNRF 125). K.W. was supported by Japan Society for the Promotion of Science (JSPS) KAKENHI grants JP19K23927 and JP20K07578. P.G. is funded by a Lundbeck postdoctoral fellowship (R322-2019-2577). A.M.-M. was funded by the Danish Cancer Society (R302-A17590). D.H.L. and Z.G. were supported by the Danish Cancer Society, Danish Cancer Society-Knæk Cancer (R311-A18224), the Independent Research Fund Denmark (8045-00057A), and the Novo Nordisk Foundation (NNF18OC0052647). Work in the lab of L.H.G. is funded by a Hallas-Møller emerging investigator grant from the Novo Nordisk Foundation (NNF20OC0059959) and a Sapere Aude grant from Independent Research Fund Denmark (0165-00092B). Q.T. is funded by a Lundbeck postdoctoral fellowship (R380-2021-1284). G.P. Z.G. H.H.G. D.G.-C. M.H.L. and P.G.: cell culture and manipulations, in situ transcription assay, EdU Click-iT assay, immunofluorescence analysis, immunoblots, qPCR, QIBC analysis, fluorescence-activated cell sorting (FACS), live cell imaging experiments combined with UVA laser irradiation, PALM microirradiation, survival assays, biochemical assays (co-immunoprecipitation and chromatin fractionation); G.P. S.H.N.M. D.C.K. and P.O.-C: cDNA library preparation and bioinformatic analysis of nanopore RNA sequencing data and interpretation; Q.T. and L.H.G.: preparation and performance of DRB/TTchem-seq, analysis and interpretation of the data acquired from the sequencing; G.P. S.H.N.M. K.W. D.H.L. L.H.G. A.M.-M. P.G. and J.B.: data analysis and interpretation and manuscript preparation. G.P. A.M.-M. P.G. and J.B.: experimental design, supervision and manuscript writing with input from all co-authors. The authors declare no competing interests.
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© 2023 The Authors
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