A global analysis of the reconstitution of PTEN function by translational readthrough of PTEN pathogenic premature termination codons

2021 ◽  
Vol 42 (5) ◽  
pp. 551-566
Author(s):  
Sandra Luna ◽  
Leire Torices ◽  
Janire Mingo ◽  
Laura Amo ◽  
Isabel Rodríguez‐Escudero ◽  
...  
Keyword(s):  
Premature Termination ◽  
Global Analysis ◽  
Premature Termination Codons ◽  
Translational Readthrough

scholarly journals Identification of Compounds That Promote Readthrough of Premature Termination Codons in the CFTR

2020 ◽  
pp. 247255522096200
Author(s):  
Emery Smith ◽  
Danijela Dukovski ◽  
Justin Shumate ◽  
Louis Scampavia ◽  
John P. Miller ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
Membrane Potential ◽  
Small Molecule ◽  
High Throughput ◽  
Premature Termination ◽  
Target Area ◽  
Mrna Levels ◽  
Premature Termination Codons ◽  
Translational Readthrough ◽  
Cftr Protein

Cystic fibrosis (CF) is caused by a mutation of the Cystic Fibrosis Transmembrane Conductance Regulator ( CFTR) gene, which disrupts an ion channel involved in hydration maintenance via anion homeostasis. Nearly 5% of CF patients possess one or more copies of the G542X allele, which results in a stop codon at residue 542, preventing full-length CFTR protein synthesis. Identifying small-molecule modulators of mutant CFTR biosynthesis that affect the readthrough of this and other premature termination codons to synthesize a fully functional CFTR protein represents a novel target area of drug discovery. We describe the implementation and integration for large-scale screening of a homogeneous, 1536-well functional G542X-CFTR readthrough assay. The assay uses HEK 293 cells engineered to overexpress the G542X-CFTR mutant, whose functional activity is monitored with a membrane potential dye. Cells are co-incubated with a CFTR amplifier and CFTR corrector to maximize mRNA levels and trafficking of CFTR to the cell surface. Compounds that allow translational readthrough and synthesis of functional CFTR chloride channels are reflected by changes in membrane potential in response to cAMP stimulation with forskolin and CFTR channel potentiation with genistein. Assay statistics yielded Z′ values of 0.69 ± 0.06. As further evidence of its suitability for high-throughput screening, we completed automated screening of approximately 666,000 compounds, identifying 7761 initial hits. Following secondary and tertiary assays, we identified 188 confirmed hit compounds with low and submicromolar potencies. Thus, this approach takes advantage of a phenotypic screen with high-throughput scalability to identify new small-molecule G542X-CFTR readthrough modulators.

scholarly journals MMADHC premature termination codons in the pathogenesis of cobalamin D disorder: Potential of translational readthrough reconstitution

2021 ◽  
Vol 26 ◽  
pp. 100710
Author(s):  
Leire Torices ◽  
Javier de las Heras ◽  
Juan Carlos Arango-Lasprilla ◽  
Jesús M. Cortés ◽  
Caroline E. Nunes-Xavier ◽  
...  
Keyword(s):  
Premature Termination ◽  
Premature Termination Codons ◽  
Translational Readthrough ◽  
Disorder Potential

scholarly journals Tobramycin is a suppressor of premature termination codons

2013 ◽  
Vol 12 (6) ◽  
pp. 806-811 ◽  
Author(s):  
Nicola Altamura ◽  
Rosa Castaldo ◽  
Alessia Finotti ◽  
Giulia Breveglieri ◽  
Francesca Salvatori ◽  
...  
Keyword(s):  
Premature Termination ◽  
Premature Termination Codons

Premature termination codons destabilize ferredoxin-1 mRNA when ferredoxin-1 is translated

2000 ◽  
Vol 21 (6) ◽  
pp. 563-569 ◽  
Author(s):  
Marie E. Petracek ◽  
Tuyen Nuygen ◽  
William F. Thompson ◽  
Lynn F. Dickey
Keyword(s):  
Premature Termination ◽  
Premature Termination Codons
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