A QPCR-based reporter system to study post-transcriptional regulation via the 3′ untranslated region of mRNA inSaccharomyces cerevisiae

Yeast ◽  
2009 ◽  
Vol 26 (7) ◽  
pp. 407-413 ◽  
Author(s):  
Kristina Lind ◽  
Joakim Norbeck
Keyword(s):  
Transcriptional Regulation ◽  
Untranslated Region ◽  
Reporter System ◽  
Post Transcriptional Regulation

Post-transcriptional regulation of ULBP1 ligand for the activating immunoreceptor NKG2D involves 3′ untranslated region

2011 ◽  
Vol 72 (6) ◽  
pp. 470-478 ◽  
Author(s):  
Heike Himmelreich ◽  
Arina Mathys ◽  
Aleksandra Wodnar-Filipowicz ◽  
Christian P. Kalberer
Keyword(s):  
Transcriptional Regulation ◽  
Untranslated Region ◽  
Post Transcriptional Regulation

scholarly journals The 3′-untranslated region of the mouse cholesterol 7α-hydroxylase mRNA contains elements responsive to post-transcriptional regulation by bile acids

1997 ◽  
Vol 328 (2) ◽  
pp. 393-399 ◽  
Author(s):  
B. Luis AGELLON ◽  
K. Sukhinder CHEEMA
Keyword(s):  
Transcriptional Regulation ◽  
Bile Acids ◽  
Untranslated Region ◽  
Transgene Expression ◽  
Hepatoma Cells ◽  
Early Gene ◽  
Transcriptional Level ◽  
Acid Uptake ◽  
Post Transcriptional Regulation ◽  
Conjugated Bile Acids

To investigate the importance of the 3ʹ-untranslated region (UTR) of the mouse cholesterol 7α-hydroxylase (cyp7) mRNA in post-transcriptional regulation of expression of the cyp7 gene, chimaeric genes encoding mRNA containing the structural sequence of chloramphenicol acetyltransferase (CAT) linked to either the 3ʹ-UTR of the mouse cyp7 mRNA or the SV40 early gene mRNA were constructed. The human cytomegalovirus (CMV) promoter was used to drive the expression of all the chimaeric genes. Thus the transgenes had identical sequences in the promoter, the regions encoding the 5ʹ-UTR and translated sequence but differed in the region encoding the 3ʹ-UTR of their respective mRNA species. The transgene containing the entire cyp7 3ʹ-UTR (designated CMV.CAT.CYP7) gave rise to CAT activity in transfected hepatoma cells that was one-quarter of that obtained in cells transfected with the transgene containing the SV40 3ʹ-UTR (designated CMV.CAT.SV40). The 3ʹ-UTR of the cyp7 mRNA contains sequences resembling AU-rich elements (AREs). Deleting eight of nine putative AREs from the CYP7 3ʹ-UTR sequence increased the CAT activity to a level greater than that observed for CMV.CAT.SV40, whereas deletion of the intron region had no effect. These results show that the AREs of the 3ʹ-UTR of the cyp7 mRNA decrease transgene expression. Bile acids are known to repress the expression of the cyp7 gene. To test whether the 3ʹ-UTR of the cyp7 mRNA has a role in this process, the expression of the chimaeric genes was evaluated in hepatoma cells competent for bile acid uptake. Conjugated bile acids, but not unconjugated bile acids, further decreased the expression of the CMV.CAT.CYP7 transgene. The same bile acids had no effect on the expression of the CMV.CAT.SV40 transgene. Deletion of the intron from the cyp7 sequence did not alter the CAT activity compared with the parental plasmid, and also did not alter the sensitivity of the transgene to the conjugated bile acids. Deletion of the AREs from the cyp7 3ʹ-UTR, which increased the expression of the transgene, did not abolish the sensitivity of the transgene to repression by conjugated bile acids. Thus the 3ʹ-UTR of the mouse cyp7 mRNA also contains elements that facilitate the further repression of transgene expression in the presence of conjugated bile acids. The results indicate that the 3ʹ-UTR of the mouse cyp7 mRNA contains information specifying regulation at the post-transcriptional level.

scholarly journals Post-transcriptional Regulation of RNase-L Expression Is Mediated by the 3′-Untranslated Region of Its mRNA

2007 ◽  
Vol 282 (11) ◽  
pp. 7950-7960 ◽  
Author(s):  
Xiao-Ling Li ◽  
Jesper B. Andersen ◽  
Heather J. Ezelle ◽  
Gerald M. Wilson ◽  
Bret A. Hassel
Keyword(s):  
Transcriptional Regulation ◽  
Untranslated Region ◽  
Rnase L ◽  
Post Transcriptional Regulation

Post-transcriptional regulation of Xwnt-8 expression is required for normal myogenesis during vertebrate embryonic development

Development ◽  
1999 ◽  
Vol 126 (15) ◽  
pp. 3371-3380
Author(s):  
Q. Tian ◽  
T. Nakayama ◽  
M.P. Dixon ◽  
J.L. Christian
Keyword(s):  
Transcriptional Regulation ◽  
Untranslated Region ◽  
Critical Role ◽  
Regulatory Elements ◽  
Rna Degradation ◽  
Transcriptional Level ◽  
Embryonic Patterning ◽  
Sequence Element ◽  
Zygotic Gene ◽  
Post Transcriptional Regulation

The Xenopus Wnt-8 gene is transiently expressed in ventral and lateral mesoderm during gastrulation and plays a critical role in patterning these tissues. In the current study, we show that the spatial and temporal pattern of expression of endogenous Xwnt-8 is regulated, in part, at a post-transcriptional level. We have identified a novel sequence element in the 3′ untranslated region of the Xwnt-8 RNA that controls the polyadenylation status of reporter and endogenous Xwnt-8 RNAs, directs rapid RNA degradation beginning precisely at the early gastrula stage, and represses translation of transcripts throughout development. Expression of endogenous Xwnt-8 is normally downregulated within lateral (presomitic) mesoderm following gastrulation. We demonstrate that rapid degradation of Xwnt-8 transcripts, mediated by these regulatory elements in the 3′ untranslated region, is essential to this process and that downregulation is required to prevent overcommitment of somitic cells to a myogenic fate. These studies demonstrate a role for post-transcriptional regulation of zygotic gene expression in vertebrate embryonic patterning.

scholarly journals The Role of the 3' Untranslated Region in the Post-Transcriptional Regulation of KLF6 Gene Expression in Hepatocellular Carcinoma

Cancers ◽  
2013 ◽  
Vol 6 (1) ◽  
pp. 28-41 ◽  
Author(s):  
Thoria Diab ◽  
Naima Hanoun ◽  
Christophe Bureau ◽  
Camille Christol ◽  
Louis Buscail ◽  
...  
Keyword(s):  
Gene Expression ◽  
Hepatocellular Carcinoma ◽  
Transcriptional Regulation ◽  
Untranslated Region ◽  
Post Transcriptional Regulation

Analysis of post-transcriptional regulation using the FunREG method

2010 ◽  
Vol 38 (6) ◽  
pp. 1608-1614 ◽  
Author(s):  
Benoît Laloo ◽  
Marion Maurel ◽  
Sandra Jalvy-Delvaille ◽  
Francis Sagliocco ◽  
Christophe F. Grosset
Keyword(s):  
Transcriptional Regulation ◽  
Translation Efficiency ◽  
Histone Deacetylase 6 ◽  
Reporter System ◽  
Rna Sequences ◽  
Experimental Conditions ◽  
Fluorescent Reporter ◽  
Wide Range ◽  
Transcriptional Deregulation ◽  
Post Transcriptional Regulation

An increasing number of arguments, including altered microRNA expression, support the idea that post-transcriptional deregulation participates in gene disturbances found in diseased tissues. To evaluate this hypothesis, we developed a method which facilitates post-transcriptional investigations in a wide range of human cells and experimental conditions. This method, called FunREG (functional, integrated and quantitative method to measure post-transcriptional regulation), connects lentiviral transduction with a fluorescent reporter system and quantitative PCR. Using FunREG, we efficiently measured post-transcriptional regulation mediated either by selected RNA sequences or regulatory factors (microRNAs), and then evaluated the contribution of mRNA decay and translation efficiency in the observed regulation. We demonstrated the existence of gene-specific post-transcriptional deregulation in liver tumour cells, and also reported a molecular link between a transcript variant abrogating HDAC6 (histone deacetylase 6) regulation by miR-433 and a rare familial genetic disease. Because FunREG is sensitive, quantitative and easy to use, many applications can be envisioned in fundamental and pathophysiological research.

scholarly journals Post-Transcriptional Regulation of the GAP-43 Gene by Specific Sequences in the 3′ Untranslated Region of the mRNA

1997 ◽  
Vol 17 (6) ◽  
pp. 1950-1958 ◽  
Author(s):  
Kao-Chung Tsai ◽  
Victor V. Cansino ◽  
Douglas T. Kohn ◽  
Rachael L. Neve ◽  
Nora I. Perrone-Bizzozero
Keyword(s):  
Transcriptional Regulation ◽  
Untranslated Region ◽  
Post Transcriptional Regulation ◽  
Specific Sequences

scholarly journals 3′-untranslated sequences mediate post-transcriptional regulation of 3-hydroxy-3-methylglutaryl-CoA reductase mRNA by 25-hydroxycholesterol

1995 ◽  
Vol 307 (1) ◽  
pp. 233-238 ◽  
Author(s):  
J W Choi ◽  
D M Peffley
Keyword(s):  
Protein Synthesis ◽  
Transcriptional Regulation ◽  
Untranslated Region ◽  
Simian Virus 40 ◽  
Beta Globin ◽  
Hmg Coa Reductase ◽  
Reductase Mrna ◽  
Coa Reductase ◽  
Post Transcriptional Regulation ◽  
Rna Levels

In an earlier study [Choi, Lundquist and Peffley (1993) Biochem. J. 296, 859-866], we determined that 25-hydroxycholesterol regulates 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase mRNA through a post-transcriptional mechanism that requires protein synthesis. To investigate whether 3′-untranslated sequences play a role in 25-hydroxycholesterol-mediated post-transcriptional control, we ligated approx. 1400 bp of the 3′-untranslated region of HMG-CoA reductase cDNA to the coding region of human beta-globin DNA. beta-Globin-3′-untranslated reductase fusion constructs were then transiently expressed in Chinese hamster ovary fibroblasts under conditions known to regulate reductase mRNA. There were no differences in beta-globin RNA levels in transfected cells incubated with or without lovastatin, a competitive inhibitor of reductase. However, in the presence of lovastatin and an oxysterol, 25-hydroxycholesterol, beta-globin RNA levels were decreased approx. 2-fold. Inhibition of protein synthesis with cycloheximide blocked the effects of 25-hydroxycholesterol on beta-globin RNA. Moreover, replacing the 3′-untranslated sequences with 1367 bp of the simian virus 40 enhancer region eliminated the regulatory effect of 25-hydroxycholesterol. Because the fusion construct has no sterol regulatory elements necessary for transcription, our results indicate that the change in beta-globin RNA occurred at a post-transcriptional level. In addition, we have shown that the 3′-untranslated region of HMG-CoA reductase cDNA imparted oxysterol-mediated post-transcriptional regulation to beta-globin RNA, an effect that required protein synthesis.

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