The intron-encoded U16 small nucleolar RNA (snoRNA) is a component of a new family of molecules which originate by processing of pre-mRNA in which they are contained. The mechanism of U16 snoRNA biosynthesis involves an initial step of endonucleolytic cleavage of the pre-mRNA with the release of a pre-snoRNA molecule; the subsequent step consists of exonucleolytic trimming that produces mature U16 molecules. In order to identify the molecular components involved in this peculiar biosynthetic pathway, we have undertaken the characterization of the endonucleolytic activity by biochemical fractionation of Xenopus laevis oocyte nuclear extract. In this paper we show the production of a protein fraction (BSF) which is highly enriched for a specific endonucleolytic activity that exactly reproduces the cleavage pattern of the U16-containing pre-mRNA identified in vivo in X. laevis oocytes and in unfractionated nuclear extract.
Download full-text PDF |
Source |
---|---|
http://dx.doi.org/10.1006/bbrc.1997.6487 | DOI Listing |
Mol Ther
September 2017
Department of Core Antisense Research, Ionis Pharmaceutics, Inc., Carlsbad, CA 92010, USA.
RNase H1-dependent antisense oligonucleotides (ASOs) are active in reducing levels of both cytoplasmic mRNAs and nuclear retained RNAs. Although ASO activity in the nucleus has been well demonstrated, the cytoplasmic activity of ASOs is less clear. Using kinetic and subcellular fractionation studies, we evaluated ASO activity in the cytoplasm.
View Article and Find Full Text PDFMol Ther
June 2008
Division of Molecular Biology, Beckman Research Institute of The City of Hope, Duarte, California 91010, USA.
Hammerhead ribozymes have been shown to silence human immunodeficiency virus-1 (HIV-1) gene expression by site-specific cleavage of viral mRNA. The two major factors that determine whether ribozymes will be effective for post-transcriptional gene silencing are colocalization of the ribozyme and the target RNAs, and the choice of an appropriate target site on the mRNA. An effective screening strategy for potential targets on the viral genome is the use of ribozyme libraries in cell culture.
View Article and Find Full Text PDFRNA Biol
October 2006
INSERM U 869, Institut Européen de Chimie et Biologie, Pessac, France.
An anti-TAR RNA aptamer called R06, which binds tightly and specifically to the trans-activation responsive (TAR) element of the human immunodeficiency virus type 1 (HIV-1) through loop-loop interactions has been previously selected.(1) We used HIV-based retroviral vectors to express the R06 aptamer. Its synthesis was driven by the U16 snoRNA.
View Article and Find Full Text PDFJ Mol Biol
November 2004
Institute Pasteur Cenci-Bolognetti, Department of Genetics and Molecular Biology, University "La Sapienza" P.le A. Moro 5, 00185 Rome, Italy.
In vertebrates almost all snoRNAs are encoded in introns of a specific subclass of polII transcripts: the TOP genes. The majority of these RNAs originate through debranching of the spliced introns, the rest through endonucleolytic cleavage of the precursor that contains them. In both cases it has been suggested that snoRNP factors associate at early steps during transcription and control snoRNA biogenesis.
View Article and Find Full Text PDFMol Ther
August 2003
Division of Molecular Biology, Beckman Research Institute of the City of Hope, Duarte, California 91010, USA.
A primary advantage of lentiviral vectors is their ability to pass through the nuclear envelope into the cell nucleus thereby allowing transduction of nondividing cells. Using HIV-based lentiviral vectors, we delivered an anti-CCR5 ribozyme (CCR5RZ), a nucleolar localizing TAR RNA decoy, or Pol III-expressed siRNA genes into cultured and primary cells. The CCR5RZ is driven by the adenoviral VA1 Pol III promoter, while the human U6 snRNA Pol III-transcribed TAR decoy is embedded in a U16 snoRNA (designated U16TAR), and the siRNAs were expressed from the human U6 Pol III promoter.
View Article and Find Full Text PDFEnter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!