Gromak, NataliaDienstbier, MartinMacias, SaraPlass, MireyaEyras, EduardoCaceres, Javier F.Proudfoot, Nicholas J.2025-06-112025-06-112211-1247WOS:000331153100003PubMed:24360955ORCID:/0000-0003-0793-6218/work/162948920https://www.webofscience.com/api/gateway?GWVersion=2&SrcApp=anu_research_portal_plus2&SrcAuth=WosAPI&KeyUT=WOS:000331153100003&DestLinkType=FullRecord&DestApp=WOS_CPLhttps://hdl.handle.net/1885/733758584Drosha is the main RNase III-like enzyme involved in the process of microRNA (miRNA) biogenesis in the nucleus. Using whole-genome ChIP-on-chip analysis, we demonstrate that, in addition to miRNA sequences, Drosha specifically binds promoterproximal regions of many human genes in a transcription-dependent manner. This binding is not associated with miRNA production or RNA cleavage. Drosha knockdown in HeLa cells downregulated nascent gene transcription, resulting in a reduction of polyadenylated mRNA produced from these gene regions. Furthermore, we show that this function of Drosha is dependent on its N-terminal protein-interaction domain, which associates with the RNA-binding protein CBP80 and RNA Polymerase II. Consequently, we uncover a previously unsuspected RNA cleavage-independent function of Drosha in the regulation of human gene expression.We thank N. Kim for providing pCK-Flag WT, E110aQ, and Delta N390 Drosha constructs, M. de Gobbi and D. Higgs for the help with ChIP-on-chip experiments, and Natalie Braun for the help with Figure S3C. The work was supported by a Wellcome Trust Programme Grant to N.J.P. and Royal Society University Research fellowship and MRC NIRG MR/J007870/1 to N.G. J.F.C. and S.M. were supported by the Medical Research Council and Wellcome Trust (grant 095518/Z/11/Z). E.E. was supported by grants from the Spanish Ministry of Science and by the Sandra Ibarra Foundation (BIO2008-01091, BIO2011-23920, and CSD2009-00080). M.P. is supported by the Novo Nordisk Foundation.12enMicrorna biogenesisBinding-proteinNuclear exportTranscriptionComplexMirnaMicroprocessorInterferenceRecognitionTermination201310.1016/j.celrep.2013.11.03284890993040