Inhibitory effect of SPE-39 due to tyrosine phosphorylation and ubiquitination on the function of Vps33B in the EGF-stimulated cells

Abstract Although SPE-39 is a binding protein to Vps33B that is one of the subunit in the mammalian HOPS complex, the elements of SPE-39 function remain unknown. Here, we show that tyrosine phosphorylation of SPE-39 following EGF stimulation plays a role in the stability of SPE-39 itself. Ubiquitination of the C-terminal region of SPE-39 was also elevated in response to EGF stimulation, and this process was regulated by the phosphorylation of Tyr-11 in SPE-39. However, association of Vps33B with SPE-39 inhibited the elevation of ubiquitination of SPE-39 following EGF stimulation, which might be responsible for the stabilization of SPE-39. Furthermore, an opposing functional relationship between SPE-39 and Vps33B on the downregulation of the EGF receptor was observed in EGF-stimulated COS-7 cells. Structured summary of protein interactions Vps33B and EGFR colocalize by fluorescence microscopy ( View interaction ) Vps33B and SPE-39 physically interact by anti bait coimmunoprecipitation ( View interaction ) Vps33B and SPE-39 physically interact by anti tag coimmunoprecipitation ( View interaction ) Vps33B and SPE-39 colocalize by fluorescence microscopy ( View interaction )