The Human DiGeorge Syndrome Critical Region Gene 8 and Its D. melanogaster Homolog Are Required for miRNA Biogenesis
Copyright policy )The Human DiGeorge Syndrome Critical Region Gene 8 and Its D. melanogaster Homolog Are Required for miRNA Biogenesis
MicroRNAs (miRNAs) represent a family of small noncoding RNAs that are found in plants and animals (for recent reviews, see ). miRNAs are expressed in a developmentally and tissue-specific manner and regulate the translational efficiency and stability of partial or fully sequence-complementary mRNAs. miRNAs are excised in a stepwise process from double-stranded RNA precursors that are embedded in long RNA polymerase II primary transcripts (pri-miRNA). Drosha RNase III catalyzes the first excision event, the release in the nucleus of a hairpin RNA (pre-miRNA), which is followed by export of the pre-miRNA to the cytoplasm and further processing by Dicer to mature miRNAs. Here, we characterize the human DGCR8, the DiGeorge syndrome critical region gene 8, and its Drosophila melanogaster homolog. We provide biochemical and cell-based readouts to demonstrate the requirement of DGCR8 for the maturation of miRNA primary transcripts. RNAi knockdown experiments of fly and human DGCR8 resulted in accumulation of pri-miRNAs and reduction of pre-miRNAs and mature miRNAs. Our results suggest that DGCR8 and Drosha interact in human cells and reside in a functional pri-miRNA processing complex.
- Rockefeller University United States
- Max Planck Society Germany
Ribonuclease III, Agricultural and Biological Sciences(all), Biochemistry, Genetics and Molecular Biology(all), Reverse Transcriptase Polymerase Chain Reaction, Blotting, Western, Oligonucleotides, Proteins, RNA-Binding Proteins, Blotting, Northern, Protein Structure, Tertiary, MicroRNAs, Drosophila melanogaster, Gene Components, Gene Expression Regulation, Animals, Drosophila Proteins, Humans, RNA Interference, RNA Polymerase II, RNA, Messenger, HeLa Cells, Plasmids
Ribonuclease III, Agricultural and Biological Sciences(all), Biochemistry, Genetics and Molecular Biology(all), Reverse Transcriptase Polymerase Chain Reaction, Blotting, Western, Oligonucleotides, Proteins, RNA-Binding Proteins, Blotting, Northern, Protein Structure, Tertiary, MicroRNAs, Drosophila melanogaster, Gene Components, Gene Expression Regulation, Animals, Drosophila Proteins, Humans, RNA Interference, RNA Polymerase II, RNA, Messenger, HeLa Cells, Plasmids
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