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PMID: 19461583 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Promoter-specific transcriptional interference and c-myc gene silencing by siRNAs in human cells.

The EMBO journal ·Vol. 28 ·No. 12 ·2009-06-17 ·Pages 1708-19

Napoli S, Pastori C, Magistri M, Carbone GM, Catapano CV

Abstract

Small interfering RNAs (siRNAs) directed to gene promoters can silence genes at the transcriptional level. siRNA-directed transcriptional silencing (RdTS) was first described in plants and yeasts and more recently in mammalian cells. RdTS has been associated with the induction of epigenetic changes and the formation of complexes containing RNA interference and chromatin-remodelling factors. Here, we show that a promoter-targeted siRNA inhibits transcription of the c-myc gene. Transcriptional silencing of c-myc did not involve changes of known epigenetic marks. Instead, the c-myc promoter-targeted siRNA interfered with transcription initiation blocking the assembly of the pre-initiation complex. Transcriptional interference depended on Argonaute 2 and a noncoding promoter-associated RNA initiated upstream and overlapping the transcription start site. Silencing of c-myc led to growth arrest, reduced clonogenic potential and senescence of c-myc over-expressing prostate cancer cells with minimal effect on normal cells. RNA-directed transcriptional interference may be a natural mechanism of transcriptional control and siRNAs targeting noncoding RNAs participating in this regulatory pathway could be valuable tools to control expression of deregulated genes in human diseases.

MeSH Terms
Argonaute Proteins Cell Line, Tumor Cell Proliferation Cellular Senescence Eukaryotic Initiation Factor-2/metabolism Gene Expression Regulation, Neoplastic Gene Knockdown Techniques Humans Promoter Regions, Genetic/genetics Proto-Oncogene Proteins c-myc/genetics RNA Interference RNA, Messenger/genetics,metabolism RNA, Neoplasm/metabolism RNA, Small Interfering/metabolism RNA, Untranslated/genetics Transcription, Genetic Tumor Stem Cell Assay
Chemicals
AGO2 protein, human Argonaute Proteins Eukaryotic Initiation Factor-2 Proto-Oncogene Proteins c-myc RNA, Messenger RNA, Neoplasm RNA, Small Interfering RNA, Untranslated
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Napoli Sara
Laboratory of Experimental Oncology, Oncology Institute of Southern Switzerland, Bellinzona, Switzerland.
Pastori Chiara
Magistri Marco
Carbone Giuseppina M
Catapano Carlo V
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
1460-2075
DOI
10.1038/emboj.2009.139
Published
2009-06-17
Epub
2009-00-21
Pages
1708-19
Language
English
Region
England
NLM ID
8208664
PMCID
PMC2699364
Subset
IM
External Links
PubMed source DOI: 10.1038/emboj.2009.139 PMC Full Text View journal details
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