Abstract 2454: Targeting the translational enhancer complex MCT-1:DenR to disrupt NHL survival

2015 ◽  
Author(s):  
Sean D. Stowe ◽  
Michael C. Cavalier ◽  
Raquel Godoy-Ruiz ◽  
Kristen J. Varney ◽  
Paul T. Wilder ◽  
...  
Keyword(s):  
Translational Enhancer

scholarly journals Ribosomal shunting mediated by a translational enhancer element that base pairs to 18S rRNA

2006 ◽  
Vol 103 (25) ◽  
pp. 9488-9493 ◽  
Author(s):  
S. A. Chappell ◽  
J. Dresios ◽  
G. M. Edelman ◽  
V. P. Mauro
Keyword(s):  
18S Rrna ◽  
Base Pairs ◽  
Enhancer Element ◽  
Translational Enhancer

The Epsilon Translational Enhancer

1991 ◽  
pp. 17-22
Author(s):  
Peter O. Olins ◽  
Catherine S. Devine ◽  
Shaukat H. Rangwala
Keyword(s):  
Translational Enhancer

scholarly journals Gene targeting in Arabidopsis via an all‐in‐one strategy that uses a translational enhancer to aid Cas9 expression

2019 ◽  
Vol 18 (4) ◽  
pp. 892-894 ◽  
Author(s):  
Fangnan Peng ◽  
Wenxin Zhang ◽  
Wenjie Zeng ◽  
Jian‐Kang Zhu ◽  
Daisuke Miki
Keyword(s):  
Gene Targeting ◽  
Translational Enhancer

The A312L 5′-UTR of Chlorella virus PBCV-1 is a translational enhancer in Arabidopsis thaliana

2009 ◽  
Vol 140 (1-2) ◽  
pp. 138-146 ◽  
Author(s):  
Phuong-son Nguyen ◽  
Deane L. Falcone ◽  
Michael V. Graves
Keyword(s):  
Arabidopsis Thaliana ◽  
Chlorella Virus ◽  
Translational Enhancer

scholarly journals Identification of an AU-rich Translational Enhancer within the Escherichia coli fepB Leader RNA

2007 ◽  
Vol 189 (11) ◽  
pp. 4028-4037 ◽  
Author(s):  
India G. Hook-Barnard ◽  
Timothy J. Brickman ◽  
Mark A. McIntosh
Keyword(s):  
Escherichia Coli ◽  
Posttranscriptional Regulation ◽  
Selective Advantage ◽  
In Vitro Transcription ◽  
Start Codon ◽  
Translational Efficiency ◽  
Rnase Protection ◽  
Ribosome Binding ◽  
Translational Enhancer

ABSTRACT The fepB gene encodes a periplasmic binding protein that is essential for the uptake of ferric enterobactin by Escherichia coli. Its transcription is regulated in response to iron levels by the Fur repressor. The fepB transcript includes a 217-nucleotide leader sequence with several features suggestive of posttranscriptional regulation. To investigate the fepB leader for its contribution to fepB expression, defined deletions and substitution mutations in the leader were characterized using fepB-phoA translational fusions. The fepB leader was found to be necessary for maximal fepB expression, primarily due to the influence of an AU-rich translational enhancer (TE) located 5′ to the Shine-Dalgarno sequence. Deletions or substitutions within the TE sequence decreased fepB-phoA expression fivefold. RNase protection and in vitro transcription-translation assays demonstrated that the TE augmented translational efficiency, as well as RNA levels. Moreover, primer extension inhibition assays showed that the TE increases ribosome binding. In contrast to the enhancing effect of the TE, the natural fepB GUG start codon decreased ribosome binding and reduced fepB expression 2.5-fold compared with the results obtained with leaders bearing an AUG initiation codon. Thus, the TE-GUG organization in fepB results in an intermediate level of expression compared to the level with AUG, with or without the TE. Furthermore, we found that the TE-GUG sequence is conserved among the eight gram-negative strains examined that have fepB genes, suggesting that this organization may provide a selective advantage.

scholarly journals Translation initiation from sequence variants of the bacteriophage T7 g10RBS in Escherichia coli and Agrobacterium fabrum

2021 ◽  
Author(s):  
Alex B. Benedict ◽  
Joshua D. Chamberlain ◽  
Diana G. Calvopina ◽  
Joel S. Griffitts
Keyword(s):  
Escherichia Coli ◽  
Recombinant Proteins ◽  
Bacteriophage T7 ◽  
Translation Efficiency ◽  
Nucleotide Substitutions ◽  
Reporter Protein ◽  
E Coli ◽  
Translational Enhancer ◽  
Multiple Promoter ◽  
Derivatives Of

Abstract Background The bacteriophage T7 gene 10 ribosome binding site (g10RBS) has long been used for robust expression of recombinant proteins in Escherichia coli. This RBS consists of a Shine–Dalgarno (SD) sequence augmented by an upstream translational “enhancer” (Enh) element, supporting protein production at many times the level seen with simple synthetic SD-containing sequences. The objective of this study was to dissect the g10RBS to identify simpler derivatives that exhibit much of the original translation efficiency. Methods and results Twenty derivatives of g10RBS were tested using multiple promoter/reporter gene contexts. We have identified one derivative (which we call “CON_G”) that maintains 100% activity in E. coli and is 33% shorter. Further minimization of CON_G results in variants that lose only modest amounts of activity. Certain nucleotide substitutions in the spacer region between the SD sequence and initiation codon show strong decreases in translation. When testing these 20 derivatives in the alphaproteobacterium Agrobacterium fabrum, most supported strong reporter protein expression that was not dependent on the Enh. Conclusions The g10RBS derivatives tested in this study display a range of observed activity, including a minimized version (CON_G) that retains 100% activity in E. coli while being 33% shorter. This high activity is evident in two different promoter/reporter sequence contexts. The array of RBS sequences presented here may be useful to researchers in need of fine-tuned expression of recombinant proteins of interest.

scholarly journals Uncoupled Expression of p33 and p92 Permits Amplification of Tomato Bushy Stunt Virus RNAs

1998 ◽  
Vol 72 (7) ◽  
pp. 5845-5851 ◽  
Author(s):  
Sara K. Oster ◽  
Baodong Wu ◽  
K. Andrew White
Keyword(s):  
Viral Genome ◽  
Stop Codon ◽  
Rna Virus ◽  
Rna Replication ◽  
Tomato Bushy Stunt Virus ◽  
Viral Rna ◽  
Rna Amplification ◽  
Viral Rnas ◽  
Reading Frame ◽  
Translational Enhancer

ABSTRACT Tomato bushy stunt virus (TBSV) is a plus-sense RNA virus which encodes a 33-kDa protein in its 5′-most open reading frame (ORF). Readthrough of the amber stop codon of the p33 ORF results in the production of a 92-kDa fusion protein. Both of these products are expressed directly from the viral genome and are suspected to be involved in viral RNA replication. We have investigated further the roles of these proteins in the amplification of viral RNAs by using a complementation system in which p33 and p92 are expressed from different viral RNAs. Our results indicate that (i) both of these proteins are necessary for viral RNA amplification; (ii) translation of these proteins can be uncoupled while maintaining amplification of viral RNAs; (iii) if compatibility requirements exist between p33 and p92, they are not exceptionally strict; and (iv) the C-terminal ∼6% of p33 is necessary for its functional activity. Interestingly, no complementation was observed when a p33-encoding replicon containing a deletion of a 3′-located segment, region 3.5, was tested. However, when 5′-capped transcripts of the same replicon were analyzed, complementation allowing for RNA amplification was observed. This ability to compensate functionally for the absence of region 3.5 by the addition of a 5′ cap suggests that this RNA segment may act as a translational enhancer for the expression of virally encoded products.

scholarly journals Two-Helper RNA System for Production of Recombinant Semliki Forest Virus Particles

1999 ◽  
Vol 73 (2) ◽  
pp. 1092-1098 ◽  
Author(s):  
C. Smerdou ◽  
P. Liljeström
Keyword(s):  
Semliki Forest Virus ◽  
Foot And Mouth Disease ◽  
Heterologous Protein Expression ◽  
Lacz Gene ◽  
Rna Recombination ◽  
Rna Molecules ◽  
Virus Particles ◽  
Translational Enhancer ◽  
Mouth Disease Virus ◽  
Spike Proteins

ABSTRACT Alphavirus expression systems based on suicidal virus particles carrying recombinant replicons have proven to be a very efficient way to deliver genes for heterologous protein expression. However, present strategies for production of such particles have biosafety limitations due to the generation, by RNA recombination, of replication-proficient viruses (RPVs). Here we describe a new packaging system for Semliki Forest virus (SFV) based on a the use of a two-helper system in which the capsid and spike proteins of the C-p62-6K-E1 polyprotein are expressed from two independent RNA molecules. The capsid gene contains a translational enhancer and therefore that sequence was also engineered in front of the spike sequence p62-6K-E1. A sequence coding for the foot-and-mouth disease virus 2A autoprotease was inserted in frame between the capsid translational enhancer and the spike genes. This allows production of the spike proteins at high levels with cotranslational removal of the enhancer sequence and normal biosynthesis of the spike complex. The autoprotease activity of the capsid protein was abolished by mutation, further increasing the biosafety of the system. Cotransfection of cells with both helper RNAs and an SFV vector replicon carrying the LacZ gene led to production of recombinant particles with titers of up to 8 × 108 particles per 106 cells. Extensive analysis failed to demonstrate the presence of any RPVs, emphasizing the high biosafety of the system based on two-helper RNAs.

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