An IMiD-induced SALL4 degron system for selective degradation of target proteins

Regulating the amount of proteins in living cells is a powerful approach for understanding the functions of the proteins. Immunomodulatory drugs (IMiDs) induce the degradation of neosubstrates by interacting with celebron (CRBN) in the cullin E3 ubiquitin ligase complex (CRL4(CRBN)). Here, we developed the IMiD-dependent Sal-like protein 4 (SALL4) degron (S4D) system for chemical protein knockdown. In transient assays, an N- or C-terminal S4D tag induced the degradation of proteins localized to various subcellular compartments, including the plasma membrane. The activity of luciferase-S4D was reduced by 90% within 3h of IMiD treatment. IMiD treatment reduced the expression of endogenous S4D-fused RelA and I kappa B alpha in knock-in (KI) experiments. Interestingly, the I kappa B alpha knockdown suggested that there may be another, unknown mechanism for RelA translocation to the nucleus. Furthermore, 5-hydroxythalidomide as a thalidomide metabolite specifically degradated S4D-tagged protein. These results indicate that the S4D system is a useful tool for cellular biology. Yamanaka et al exploit the fact that immunomodulatory drugs (IMiDs) interacts with the E3 ligase CRL4-CRBN to induce degradation of alternative substrates including Sal-like protein 4 (SALL4) to develop a degradation system that targets proteins tagged with a SALL4-like degron. This tool can degrade proteins at various cellular locations.