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Bhat, S. Y.

Publications and source records attributed to Bhat, S. Y..

2 recordsLinked to original sources

Mechanism-Based Discovery of New TRIDs Targeting Ribosome Pockets

Approximately 10% of all human pathogenic mutations are nonsense mutations which give rise to premature termination codon (PTC) diseases, leading to an estimated 35 million people worldwide who harbor such mutations. To date, no approaches leading to medical treatments which increase PTC readthrough have received FDA approval. However, one well-tolerated translation readthrough inducing drug (TRID), ataluren, was conditionally approved for 11 years by the European Medicines Agency for treatment of nonsense mutation Duchenne Muscular Dystrophy. In recent work we have shown that ataluren increases readthrough indirectly, by exclusively inhibiting termination without directly affecting readthrough by suppressor aminoacyl-tRNAs (sup-aa-tRNAs). Based on the idea that this mode of action (MoA) might be the reason for the low toxicity of ataluren, we have discovered potential new TRIDs, which target pockets within the ribosome to which the termination factor eRF1 binds but sup-aa-tRNAs do not. These new TRIDs, dipeptide derivatives of eRF1 and some Sartans, mimic the MoA of ataluren. Three Sartans, which are angiotensin II receptor blockers and FDA-approved for treatment of hypertension, are significantly more potent than ataluren in our reconstituted in vitro readthrough assay. These compounds might be more effective than ataluren in treating some PTC diseases. We anticipate that further development of these lead compounds will lead to more general effectiveness, used both alone and in combination with other classes of readthrough inducers.

biochemistry↗

Suppression of nonsense mutations by small, cyclic peptides

Premature termination codons in mRNAs result from nonsense mutations and hinder the translation of full-length, functional proteins. Nonsense mutations cause numerous serious genetic diseases, including cystic fibrosis and Duchenne muscular dystrophy. Several small-molecule drugs have been reported that could potentially ameliorate these diseases by promoting translational readthrough at the premature termination codon. However, utilization of many of these molecules faces problems such as limited efficacy or high cellular toxicity. Using a selection strategy in Saccharomyces cerevisiae coupling suppression of endogenous nonsense mutations to cell survival, we identified ten readthrough-promoting cyclic peptides from a DNA-encoded library. The selected cyclic peptides suppress nonsense mutations in various reporter genes, and the candidates inducing the highest readthrough levels display no observable cytotoxicity in yeast. Mutational analysis of the most promising cyclic peptide demonstrate that most amino acid side chains contribute to the readthrough-stimulating activity. Importantly, this cyclic peptide appears to bind directly to the eukaryotic core translation machinery and promotes readthrough in vitro by interfering with ribosomal decoding. Our results suggest that small, cyclic peptides selected in vivo could represent a novel drug type to treat the many incurable human genetic diseases that are caused by nonsense mutations.

molecular biology↗