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) |