In the last few years, aminoglycosides have been investigated to suppress premature translation termination by inducing a ribosomal read-through of premature, but not the natural, termination codons (PTCs). The functional consequences of these PTCs are minimized by the nonsense-mediated RNA decay (NMD) pathway, a cellular mechanism, aimed to detect and degrade PTC containing mRNA. Tobramycin is an aminoglycoside antibiotic, normally used to treat Pseudomonas aeruginosa pulmonary infection in cystic fibrosis (CF), a recessive genetic disease characterized by persistant pulmonary infection and extensive lung inflammation. We have analyzed the possible use of Tobramycin: (a) to alter the expression of IL-8 in CF IB3-1 cells induced with pro-inflammatory cytokine TNF- and, (b) to suppress PTCs (present in 5-10% of CF alleles) by read-through mechanism in Saccharomyces cerevisiae yeast strains carrying a dual luc reporter system separated by nonsense codon and having functional or abolished NMD mechanism by deletion of UPF1, one of the key NMD factor. The results demonstrate that Tobramycin is a good inhibitor of IL-8 expression and presents a potential read-through activity on PTCs, but not with significant effect on the NMD mechanism. It will be interesting investigate the potential read-through activity of other well-known aminoglycosides overcoming pre-clinical studies.
Effects of Tobramycin activity on premature termination codons and interleukin 8 gene expression
BREVEGLIERI, Giulia;FINOTTI, Alessia;SALVATORI, Francesca;ZUCCATO, Cristina;GAMBARI, Roberto;BORGATTI, Monica
2012
Abstract
In the last few years, aminoglycosides have been investigated to suppress premature translation termination by inducing a ribosomal read-through of premature, but not the natural, termination codons (PTCs). The functional consequences of these PTCs are minimized by the nonsense-mediated RNA decay (NMD) pathway, a cellular mechanism, aimed to detect and degrade PTC containing mRNA. Tobramycin is an aminoglycoside antibiotic, normally used to treat Pseudomonas aeruginosa pulmonary infection in cystic fibrosis (CF), a recessive genetic disease characterized by persistant pulmonary infection and extensive lung inflammation. We have analyzed the possible use of Tobramycin: (a) to alter the expression of IL-8 in CF IB3-1 cells induced with pro-inflammatory cytokine TNF- and, (b) to suppress PTCs (present in 5-10% of CF alleles) by read-through mechanism in Saccharomyces cerevisiae yeast strains carrying a dual luc reporter system separated by nonsense codon and having functional or abolished NMD mechanism by deletion of UPF1, one of the key NMD factor. The results demonstrate that Tobramycin is a good inhibitor of IL-8 expression and presents a potential read-through activity on PTCs, but not with significant effect on the NMD mechanism. It will be interesting investigate the potential read-through activity of other well-known aminoglycosides overcoming pre-clinical studies.I documenti in SFERA sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
