An RNA-directed nuclease mediates post-transcriptional gene silencing in Drosophila cells

Hammond, S. M., Bernstein, E., Beach, D., Hannon, G. J. (2000) An RNA-directed nuclease mediates post-transcriptional gene silencing in Drosophila cells. Nature, 404 (6775). pp. 293-296. ISSN 00280836 (ISSN)

URL: http://www.ncbi.nlm.nih.gov/pubmed/10749213
DOI: 10.1038/35005107

Abstract

In a diverse group of organisms that includes Caenorhabditis elegans, Drosophila, planaria, hydra, trypanosomes, fungi and plants, the introduction of double-stranded RNAs inhibits gene expression in a sequence-specific manner. These responses, called RNA interference or post-transcriptional gene silencing, may provide anti-viral defence, modulate transposition or regulate gene expression. We have taken a biochemical approach towards elucidating the mechanisms underlying this genetic phenomenon. Here we show that 'loss-of- function' phenotypes can be created in cultured Drosophila cells by transfection with specific double-stranded RNAs. This coincides with a marked reduction in the level of cognate cellular messenger RNAs. Extracts of transfected cells contain a nuclease activity that specifically degrades exogenous transcripts homologous to transfected double-stranded RNA. This enzyme contains an essential RNA component. After partial purification, the sequence-specific nuclease co-fractionates with a discrete, ~25-nucleotide RNA species which may confer specificity to the enzyme through homology to the substrate mRNAs.

Item Type: Paper
Uncontrolled Keywords: nuclease article Drosophila enzyme activity enzyme purification gene expression regulation gene silencing priority journal sequence homology Animals Antigens, CD8 Cell Cycle Cells, Cultured Cyclin E Endoribonucleases Humans Lac Operon RNA, Double-Stranded RNA, Messenger Substrate Specificity Transcription, Genetic Transfection Caenorhabditis elegans Fungi Hydra Invertebrata Planaria
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > transcription
organism description > animal > insect > Drosophila
bioinformatics > genomics and proteomics > genetics & nucleic acid processing
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > RNA regulation
CSHL Authors:
Communities: CSHL labs > Hannon lab
Depositing User: Matt Covey
Date: 2000
Date Deposited: 09 Jan 2013 17:34
Last Modified: 09 Jan 2013 17:34
Related URLs:
URI: http://repository.cshl.edu/id/eprint/26476

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