Fig. 7
MoCsn5 inhibits autophagy by suppressing the K48-ubiquitination of MoTor. A and B Colony morphology and relative growth rate of 70-15, ΔMocsn5 and Mocsn5-C on CM supplemented with 100 ng/mL rapamycin for 9 days. *p < 0.05. C The phosphorylation levels of MoRps6 in the 70-15 and ΔMocsn5 strains were detected by anti-phospho-Rps6 and anti-Rps6 antibodies. The phosphorylation levels of MoRps6 were calculated as phospho-MoRps6/MoRps6. The protein content of actin was used as a control. D Protein levels of MoTor in 70-15, ΔMocsn5 and Mocsn5-C. The strains were cultured in liquid CM at 25 ℃ for 36 h. The protein content of actin was used as a control. Gene knockout and complementation of MoCSN5 were performed in 70-15 expressing MoTor-Flag to ensure consistent transcript levels of MoTor. E Transcript levels of MoTor in 70-15 and ΔMocsn5. The error bars represent the standard deviations. F Ubiquitination levels of MoTor in 70-15, ΔMocsn5 and Mocsn5-C. After lysis, the strains were immunoprecipitated with anti-Flag beads, and a western blot analysis was subsequently performed with an anti-Flag antibody and an anti-ubiquitin antibody. The level of ubiquitinated MoTor was calculated as ubiquitin/MoTor-Flag. G K48-ubiquitination levels of MoTor in 70-15, ΔMocsn5 and Mocsn5-C. After lysis, the strains were immunoprecipitated with anti-Flag beads, and a western blot analysis was subsequently performed with an anti-Flag antibody and an anti-K48-ubiquitin antibody. The K48-ubiquitination levels of MoTor were calculated as K48-ubiquitin/MoTor-Flag