Antibody RING-Mediated Destruction of Endogenous Proteins

TY - JOUR

T1 - Antibody RING-Mediated Destruction of Endogenous Proteins

AU - Ibrahim, Adel F. M.

AU - Shen, Linnan

AU - Tatham, Michael H.

AU - Dickerson, David

AU - Prescott, Alan R.

AU - Abidi, Naima

AU - Xirodimas, Dimitris P.

AU - Hay, Ronald T.

N1 - This work was supported by Wellcome Trust Investigator Awards (098391/Z/12/Z and 217196/Z/19/Z) and Cancer Research UK Programme Grant (C434/A21747) to R.T.H. Naima Abidi was supported by a post-doctoral fellowship from Fondation Recherche Médicale

PY - 2020/7/2

Y1 - 2020/7/2

N2 - To understand gene function, the encoding DNA or mRNA transcript can be manipulated and the consequences observed. However, these approaches do not have a direct effect on the protein product of the gene, which is either permanently abrogated or depleted at a rate defined by the half-life of the protein. We therefore developed a single-component system that could induce the rapid degradation of the specific endogenous protein itself. A construct combining the RING domain of ubiquitin E3 ligase RNF4 with a protein-specific camelid nanobody mediates target destruction by the ubiquitin proteasome system, a process we describe as antibody RING-mediated destruction (ARMeD). The technique is highly specific because we observed no off-target protein destruction. Furthermore, bacterially produced nanobody-RING fusion proteins electroporated into cells induce degradation of target within minutes. With increasing availability of protein-specific nanobodies, this method will allow rapid and specific degradation of a wide range of endogenous proteins.

AB - To understand gene function, the encoding DNA or mRNA transcript can be manipulated and the consequences observed. However, these approaches do not have a direct effect on the protein product of the gene, which is either permanently abrogated or depleted at a rate defined by the half-life of the protein. We therefore developed a single-component system that could induce the rapid degradation of the specific endogenous protein itself. A construct combining the RING domain of ubiquitin E3 ligase RNF4 with a protein-specific camelid nanobody mediates target destruction by the ubiquitin proteasome system, a process we describe as antibody RING-mediated destruction (ARMeD). The technique is highly specific because we observed no off-target protein destruction. Furthermore, bacterially produced nanobody-RING fusion proteins electroporated into cells induce degradation of target within minutes. With increasing availability of protein-specific nanobodies, this method will allow rapid and specific degradation of a wide range of endogenous proteins.

KW - ARMeD

KW - E3 ligase

KW - nanobody-RING fusion

KW - proteasome

KW - protein degradation

KW - ubiquitin

UR - https://www.scopus.com/pages/publications/85086393441

U2 - 10.1016/j.molcel.2020.04.032

DO - 10.1016/j.molcel.2020.04.032

M3 - Article

C2 - 32454028

SN - 1097-2765

VL - 79

SP - 155

EP - 166

JO - Molecular Cell

JF - Molecular Cell

IS - 1

ER -