Stabilization of slow troponin C polypeptide compensates for its reduced synthesis in antisense oligodeoxynucleotide-treated cells

Abstract

The expression of genes for contractile proteins during myogenesis is coordinately regulated. Uncoupling the expression of the slow/cardiac troponin C (sTnC) gene from this process with an antisense phosphorothioate oligodeoxynucleotide (ODN) was used to examine the presence of any post-transcriptional mechanisms for regulating muscle protein synthesis. Approximately 70 and 50% decreases in sTnC polypeptide synthesis and mRNA levels, respectively, were achieved after 4 days antisense treatment. This decrease in sTnC polypeptide synthesis was not reflected in a similar decline in the steady-state level of this polypeptide. Extension of the ODN treatment to 7 days was required to produce a substantial decrease in the steady-state level of sTnC polypeptide. Our investigation suggests that during the 4-day treatment, the affected cells stabilized the sTnC polypeptide level by increasing its half-life. However, the stabilizing effect appears to be overridden during prolonged (7 days) antisense ODN treatment. Measurement of the polypeptide synthesis and mRNA levels of several contractile proteins showed no evidence of cross-regulation among the genes to coordinately regulate their expression levels.