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Series GSE140209 Query DataSets for GSE140209
Status Public on Jul 08, 2020
Title The DGCR8-UGU interaction in microRNA biogenesis
Organism Homo sapiens
Experiment type Non-coding RNA profiling by high throughput sequencing
Summary Human Microprocessor is a trimeric complex composed of the RNase III enzyme, DROSHA, and a dimer of DGCR8 (its cofactor). Microprocessor initiates the biogenesis of microRNA by processing primary microRNAs (pri-miRNAs), during which, its cleavage efficiency and accuracy are enhanced because DGCR8 interacts with the apical UGU motif of pri-miRNAs. However, the mechanism of DGCR8-UGU interaction and influence of this interaction on the expression of cellular miRNA are still elusive. In this study, we demonstrated that the Rhed domain (i.e., the RNA-binding heme domain, amino acids 285-478) of DGCR8 is sufficient to recognize and interact with UGU. In addition, we identified three amino acid residues in Rhed (amino acids 361-363), which are critical for the UGU interaction, and we showed that these residues are essential for the Microprocessor complex to accurately and efficiently process pri-miRNAs in vitro. Furthermore, we found that within the DGCR8 dimer, the UGU-binding site from just one monomer is capable of discriminating between UGU- and non-UGU-containing pri-miRNAs. Finally, we showed that these amino acids are more important for the expression of UGU miRNAs than non-UGU miRNAs in human cells. This study improves our understanding of the substrate-recognizing mechanism of DGCR8, and implicates the roles of this recognition in differentiating miRNA expression in human cells.
Overall design DGCR8-KO cells were transfected with pCK empty, pCK-DGCR8 WT or pCK-DGCR8 mut1 plasmid. Small RNA sequencing libraries of the transfected DGCR8-KO cells were generated using the TruSeq Small RNA Library Prep kit. The experiment was repeated three times.
Contributor(s) Nguyen TA, Dang TL, Li S, Nguyen TL, Nguyen TD
Citation(s) 32620823
Submission date Nov 11, 2019
Last update date Jul 08, 2020
Contact name Duc Trung Nguyen
Phone +582 93510497
Organization name The Hong Kong University of Science and Technology
Department Life Science
Lab Anh Lab
Street address Clear Water Bay
City Hong Kong
ZIP/Postal code 00000
Country Hong Kong
Platforms (1)
GPL24676 Illumina NovaSeq 6000 (Homo sapiens)
Samples (9)
GSM4568420 DGCR8 KO transfected with pCK empty plasmid replicate 1
GSM4568421 DGCR8 KO transfected with pCK empty plasmid replicate 2
GSM4568422 DGCR8 KO transfected with pCK empty plasmid replicate 3
BioProject PRJNA588809
SRA SRP229498

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Supplementary file Size Download File type/resource
GSE140209_rawcount.csv.gz 10.3 Kb (ftp)(http) CSV
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