Connexins have relative short half-lives.
Connexin 31.1 (Cx31.1) was newly reported to be down-regulated in
non-small cell lung cancer cell lines, and displayed tumour-suppressive properties. However, no reports describing how a cell regulates Cx31.1 level were found. In this study, Cx31.1 was suggested to be degraded through both
ubiquitin-
proteasome system (UPS) and autophagy. Blockage of UPS with
MG-132 increased Cx31.1 level, but could not inhibit the degradation of Cx31.1 completely. In H1299 cells stably expressing Cx31.1, Cx31.1 reduced when autophagy was induced through
starvation or
Brefeldin A treatment. Knockdown of
autophagy-related protein ATG5 could increase the cellular level of Cx31.1 both under normal growth condition and
starvation-induced autophagy. Colocalization of Cx31.1 and autophagy marker light chain 3 (LC3) was revealed by immunofluorescence analysis. Coimmunoprecipitation and immunofluorescence showed that Cx31.1 might interact with
clathrin heavy chain which was newly reported to regulate autophagic lysosome reformation (ALR) and controls lysosome homoeostasis. When
clathrin expression was knockdown by
siRNA treatment, the level of Cx31.1 increased prominently both under normal growth condition and
starvation-induced autophagy. Under
starvation-induced autophagy, LC3-II levels were slightly accumulated with siCla. treatment compared to that of
siNC, which could be ascribed to that
clathrin knockdown impaired the late stage of autophagy, ALR. Taken together, we found autophagy contributed to Cx31.1 degradation, and
clathrin might be involved in the autophagy of Cx31.1.