Ubiquitin-ligase and deubiquitinating gene expression in stretched rat skeletal muscle.

In order to gain insight into intracellular mechanisms involved in longitudinal growth of skeletal muscle, we determined gene expression of ubiquitin-ligases (MAFbx/atrogin-1, E3 alpha, and MuRF-1) and deubiquitinating enzymes (UBP45, UBP69, and USP28) at different time-points (24, 48, and 96 h) of continuous stretch of the soleus and tibialis anterior (TA) muscles. In the soleus, real-time polymerase chain reaction (PCR) showed that MAFbx/atrogin-1, E3 alpha, and MuRF-1 gene expression was downregulated, peaking at 24-48 h. Gene expression of all deubiquitinating enzymes increased with continuous stretch of soleus. In the TA, gene expression of the ubiquitin-ligases MAFbx/atrogin-1 and MuRF-1 was elevated, whereas expression of UBP45 and UBP69 was downregulated. Western blot analysis showed that the overall ubiquitination level decreased in the soleus and increased in the TA during stretch. These results suggest that ubiquitin-ligases and deubiquitinating enzymes are involved in longitudinal growth induced by continuous muscle stretch.