RRC ID 81457
Author Yamanaka S, Shoya Y, Matsuoka S, Nishida-Fukuda H, Shibata N, Sawasaki T.
Title An IMiD-induced SALL4 degron system for selective degradation of target proteins.
Journal Commun Biol
Abstract 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 (CRL4CRBN). 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 3 h of IMiD treatment. IMiD treatment reduced the expression of endogenous S4D-fused RelA and IκBα in knock-in (KI) experiments. Interestingly, the IκBα 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.
Volume 3(1)
Pages 515
Published 2020-9-18
DOI 10.1038/s42003-020-01240-5
PII 10.1038/s42003-020-01240-5
PMID 32948804
PMC PMC7501283
MeSH Cell Membrane / genetics Cell Membrane / immunology Gene Knockdown Techniques / methods HeLa Cells Humans Immunologic Factors / genetics* Immunologic Factors / immunology Immunologic Factors / pharmacology Proteolysis* Substrate Specificity Thalidomide / analogs & derivatives Thalidomide / immunology Thalidomide / metabolism* Thalidomide / pharmacology Transcription Factor RelA / genetics Transcription Factor RelA / immunology Transcription Factors / genetics* Transcription Factors / immunology Transcriptional Elongation Factors / genetics Ubiquitin-Protein Ligases / genetics* Ubiquitin-Protein Ligases / immunology
Resource
DNA material pCMV-VSV-G-RSV-Rev (RDB04393) pCAG-HIVgp (RDB04394)