scholarly journals U1 snRNA promotes the selection of nearby 5' splice sites by U6 snRNA in mammalian cells.

1996 ◽  
Vol 10 (3) ◽  
pp. 338-350 ◽  
Author(s):  
D Y Hwang ◽  
J B Cohen
Keyword(s):  
Mammalian Cells ◽  
Splice Sites ◽  
U6 Snrna ◽  
U1 Snrna ◽  
Selection Of

scholarly journals The Pre-mRNA 5′ Cap Determines Whether U6 Small Nuclear RNA Succeeds U1 Small Nuclear Ribonucleoprotein Particle at 5′ Splice Sites

1998 ◽  
Vol 18 (12) ◽  
pp. 7510-7520 ◽  
Author(s):  
Laura O’Mullane ◽  
Ian C. Eperon
Keyword(s):  
Splice Site ◽  
Major Effect ◽  
Small Nuclear Rna ◽  
Splice Sites ◽  
U6 Snrna ◽  
Small Nuclear Ribonucleoprotein ◽  
U1 Snrna ◽  
U1 Snrnp ◽  
Nuclear Rna ◽  
Nuclear Ribonucleoprotein

ABSTRACT Efficient splicing of the 5′-most intron of pre-mRNA requires a 5′ m7G(5′)ppp(5′)N cap, which has been implicated in U1 snRNP binding to 5′ splice sites. We demonstrate that the cap alters the kinetic profile of U1 snRNP binding, but its major effect is on U6 snRNA binding. With two alternative wild-type splice sites in an adenovirus pre-mRNA, the cap selectively alters U1 snRNA binding at the site to which cap-independent U1 snRNP binding is stronger and that is used predominantly in splicing; with two consensus sites, the cap acts on both, even though one is substantially preferred for splicing. However, the most striking quantitative effect of the 5′ cap is neither on U1 snRNP binding nor on the assembly of large complexes but on the replacement of U1 snRNP by U6 snRNA at the 5′ splice site. Inhibition of splicing by a cap analogue is correlated with the loss of U6 interactions at the 5′ splice site and not with any loss of U1 snRNP binding.

The effect of U1 snRNA binding free energy on the selection of 5′ splice sites

2005 ◽  
Vol 333 (1) ◽  
pp. 64-69 ◽  
Author(s):  
Jianning Bi ◽  
Huiyu Xia ◽  
Fei Li ◽  
Xuegong Zhang ◽  
Yanda Li
Keyword(s):  
Free Energy ◽  
Binding Free Energy ◽  
Splice Sites ◽  
U1 Snrna ◽  
Selection Of

A single-round selection of selective DNA aptamers for mammalian cells by polymer-enhanced capillary transient isotachophoresis

The Analyst ◽  
2017 ◽  
Vol 142 (21) ◽  
pp. 4030-4038 ◽  
Author(s):  
Kazuki Hirose ◽  
Maho Tsuchida ◽  
Hinako Asakura ◽  
Koji Wakui ◽  
Keitaro Yoshimoto ◽  
...  
Keyword(s):  
Capillary Electrophoresis ◽  
Mammalian Cells ◽  
Dna Aptamer ◽  
Dna Aptamers ◽  
Aptamer Selection ◽  
Selection For ◽  
First Time ◽  
Selection Of

A single-round DNA aptamer selection for mammalian cells was successfully achieved for the first time using a capillary electrophoresis (CE)-based methodology.

scholarly journals U1 small nuclear RNAs with altered specificity can be stably expressed in mammalian cells and promote permanent changes in pre-mRNA splicing.

1993 ◽  
Vol 13 (5) ◽  
pp. 2666-2676 ◽  
Author(s):  
J B Cohen ◽  
S D Broz ◽  
A D Levinson
Keyword(s):  
Splice Site ◽  
Mammalian Cells ◽  
Mrna Processing ◽  
Mrna Splicing ◽  
Small Nuclear Rna ◽  
Splice Sites ◽  
Gene Complementation ◽  
Small Nuclear Rnas ◽  
Nuclear Rna ◽  
Mutant Genes

Pre-mRNA 5' splice site activity depends, at least in part, on base complementarity to U1 small nuclear RNA. In transient coexpression assays, defective 5' splice sites can regain activity in the presence of U1 carrying compensatory changes, but it is unclear whether such mutant U1 RNAs can be permanently expressed in mammalian cells. We have explored this issue to determine whether U1 small nuclear RNAs with altered specificity may be of value to rescue targeted mutant genes or alter pre-mRNA processing profiles. This effort was initiated following our observation that U1 with specificity for a splice site associated with an alternative H-ras exon substantially reduced the synthesis of the potentially oncogenic p21ras protein in transient assays. We describe the development of a mammalian complementation system that selects for removal of a splicing-defective intron placed within a drug resistance gene. Complementation was observed in proportion to the degree of complementarity between transfected mutant U1 genes and different defective splice sites, and all cells selected in this manner were found to express mutant U1 RNA. In addition, these cells showed specific activation of defective splice sites presented by an unlinked reporter gene. We discuss the prospects of this approach to permanently alter the expression of targeted genes in mammalian cells.

scholarly journals Selection of Transfected Mammalian Cells

2003 ◽  
Vol 62 (1) ◽  
Author(s):  
Richard Mortensen ◽  
Jonathan D. Chesnut ◽  
James P. Hoeffler ◽  
Robert E. Kingston
Keyword(s):  
Mammalian Cells ◽  
Selection Of

scholarly journals Selection of Retroviral Reverse Transcription Primer Is Coordinated with tRNA Biogenesis

2003 ◽  
Vol 77 (16) ◽  
pp. 8695-8701 ◽  
Author(s):  
Nathan J. Kelly ◽  
Matthew T. Palmer ◽  
Casey D. Morrow
Keyword(s):  
Reverse Transcription ◽  
Nuclear Export ◽  
Mammalian Cells ◽  
Leukemia Virus ◽  
Wild Type ◽  
Primer Selection ◽  
Yeast Trna ◽  
Trna Primer ◽  
Hiv 1 ◽  
Selection Of

ABSTRACT Initiation of retrovirus reverse transcription requires the selection of a tRNA primer from the intracellular milieu. To investigate the features of primer selection, a human immunodeficiency virus type 1 (HIV-1) and a murine leukemia virus (MuLV) were created that require yeast tRNAPhe to be supplied in trans for infectivity. Wild-type yeast tRNAPhe expressed in mammalian cells was transported to the cytoplasm and aminoacylated. In contrast, tRNAPhe without the D loop (tRNAPheD−) was retained within the nucleus and did not complement infectivity of either HIV-1 or MuLV; however, infectivity was restored when tRNAPheD− was directly transfected into the cytoplasm of cells. A tRNAPhe mutant (tRNAPheUUA) that did not have the capacity to be aminoacylated was transported to the cytoplasm and did complement infectivity of both HIV-1 and MuLV, albeit at a level less than that with wild-type tRNAPhe. Collectively, our results demonstrate that the tRNA primer captured by HIV-1 and MuLV occurs after nuclear export of tRNA and supports a model in which primer selection for retroviruses is coordinated with tRNA biogenesis at the intracellular site of protein synthesis.

scholarly journals Pathways for selection of 5′ splice sites by U1 snRNPs and SF2/ASF.

1993 ◽  
Vol 12 (9) ◽  
pp. 3607-3617 ◽  
Author(s):  
I.C. Eperon ◽  
D.C. Ireland ◽  
R.A. Smith ◽  
A. Mayeda ◽  
A.R. Krainer
Keyword(s):  
Splice Sites ◽  
Selection Of

scholarly journals A new vector, based on the PolII promoter for the U1 snRNA gene, for the expression of siRNAs in mammalian cells

2004 ◽  
Vol 10 (1) ◽  
pp. 191-199 ◽  
Author(s):  
Michela Alessandra Denti ◽  
Alessandro Rosa ◽  
Olga Sthandier ◽  
Fernanda Gabriella De Angelis ◽  
Irene Bozzoni
Keyword(s):  
Mammalian Cells ◽  
Snrna Gene ◽  
U1 Snrna

Novel Method for Selection of tRNA-Driven Ribozymes with Enhanced Stability in Mammalian Cells

2002 ◽  
Vol 12 (5) ◽  
pp. 341-352 ◽  
Author(s):  
Masayuki Sano ◽  
Tomoko Kuwabara ◽  
Masaki Warashina ◽  
Akiyoshi Fukamizu ◽  
Kazunari Taira
Keyword(s):  
Mammalian Cells ◽  
Novel Method ◽  
Enhanced Stability ◽  
Selection Of

scholarly journals Selection of cryptic 5′ splice sites by group II intron RNAsin vitro

1988 ◽  
Vol 16 (15) ◽  
pp. 7383-7395 ◽  
Author(s):  
Manfred W. Müller ◽  
Rudolf J. Schweyen ◽  
Carlo Schmelzer
Keyword(s):  
Group Ii Intron ◽  
Splice Sites ◽  
Group Ii ◽  
Selection Of
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