Abstract
RNase III DROSHA is upregulated in multiple cancers and contributes to tumor progression by hitherto unclear mechanisms. Here, we demonstrate that DROSHA interacts with β-Catenin to transactivate STC1 in an RNA cleavage-independent manner, contributing to breast cancer stem-like cell (BCSC) properties. DROSHA mRNA stability is enhanced by N6-methyladenosine (m6A) modification which is activated by AURKA in BCSCs. AURKA stabilizes METTL14 by inhibiting its ubiquitylation and degradation to promote DROSHA mRNA methylation. Moreover, binding of AURKA to DROSHA transcript further strengthens the binding of the m6A reader IGF2BP2 to stabilize m6A-modified DROSHA. In addition, wild-type DROSHA, but not an m6A methylation-deficient mutant, enhances BCSC stemness maintenance, while inhibition of DROSHA m6A modification attenuates BCSC traits. Our study unveils the AURKA-induced oncogenic m6A modification as a key regulator of DROSHA in breast cancer and identifies a novel DROSHA transcriptional function in promoting the BCSC phenotype.
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Data availability
The RNA-seq and microarray data generated by this study have been deposited in the GEO database under the accession number GSE128428 (RNA-seq) and GSE125705 (microarray). Human breast tumor gene expression and clinical data were derived from TCGA Research Network (http://cancergenome.nih.gov/). Previously published meRIP-seq data and microarray data were re-analyzed. They are available under accession codes: GSE60213, GSE29714, GSE90642 and GSE54365 for meRIP-seq; GSE7513 and GSE15192 for microarray.
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Acknowledgements
This work was supported by the National Key R&D Program of China (2019YFA0110300 to Q.Liu and 2017YFA0505600-04 to Q.Liu), the National Natural Science Foundation of China (81820108024 to Q.Liu, 81630005 to Q.Liu, 81972786 to J.X., 81573025 to Q.Liu, 81703062 to L.H. and 81703091 to F.A.), Program for Changjiang Scholars and Innovative Research Team in University of Ministry of Education of China (IRT_17R15), Innovative Research Team in University of Liaoning (LT2017001 to Q.Liu), the Natural Science Foundation of Liaoning (2019-BS-081 to F.P.), the “Seedling cultivation” program for young scientific and technological talents of Liaoning (LZ2019067 to B.C. and 2020 to F.P.), the program for climbing Scholars of Liaoning, the Science and Technology Innovation Foundation of Dalian (2020JJ25CY008 to Q.Liu), Dalian High-level Talent Innovation Program (2016RD12 to Q.Liu), International Scientific and Technological Cooperation of Dalian (2015F11GH095 to Q.Liu), the Natural Science Foundation of Guangdong (2016A030311038 and 2017A030313608 to Q.Liu, 2017A020215098 to Z.W.), the Science and Technology Planning Project of Guangzhou (201804020044 to Q.Liu). E.W-F.L.’s work is supported by MRC (MR/N012097/1), CRUK (C37/A12011; C37/18784), Breast Cancer Now (2012MayPR070; 2012NovPhD016; 2014NovPhD326). The authors thank Prof. Yungui Yang (Beijing Institute of Genomics, CAS) for scientific advice and technical assistance, especially meRIP-seq data analysis and the method of meRIP-qPCR assay. The authors thank Prof. Lingqiang Zhang (Beijing Institute of Lifeomics, Beijing, China) for a gift of HA-Ub plasmid.
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Q.Liu, J.X and F.P. conceived and designed the entire project. Q.Liu, F.P., J.X., B.H. and Z.W., designed and supervised the research. F.P., B.C., Q.Liang, S.Z., H.Zou, M.L., H.Zhao, Y.M., J.H. and J.Z. performed the experimental data analyses and/or experimental planning. H.Zou, Y.L., J.L. and Z.L., performed the transcriptome-wide and meRIP-seq data analyses. B.L., S.L., J.C. and F.W. performed the proteomic analyses by LC-MS. P.C. and Z.S. performed MD simulation. F.A. drafted the working model. L.X. collected breast tumor specimens and conjugated normal breast specimens. Q.Liu, J.X., F.P., B.C. and E.W-F.L. contributed reagents/analytic tools and/or grant support. F.P., B.C., Q.Liu, J.X., B.H., Q.Liang, S.Z., Y.Z. and E.W-F.L. wrote and revised the manuscript. All authors discussed the results and commented on the manuscript.
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Peng, F., Xu, J., Cui, B. et al. Oncogenic AURKA-enhanced N6-methyladenosine modification increases DROSHA mRNA stability to transactivate STC1 in breast cancer stem-like cells. Cell Res 31, 345–361 (2021). https://doi.org/10.1038/s41422-020-00397-2
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DOI: https://doi.org/10.1038/s41422-020-00397-2
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