RRC ID 73107
Author Suzuki S, Nakamura A, Hatano Y, Yoshikawa M, Yoshii T, Sawada S, Atsuta-Tsunoda K, Aoki K, Tsukiji S.
Title A chemogenetic platform for controlling plasma membrane signaling and synthetic signal oscillation.
Journal Cell Chem Biol
Abstract Chemogenetic methods enabling the rapid translocation of specific proteins to the plasma membrane (PM) in a single protein-single ligand manner are useful tools in cell biology. We recently developed a technique, in which proteins fused to an Escherichia coli dihydrofolate reductase (eDHFR) variant carrying N-terminal hexalysine residues are recruited from the cytoplasm to the PM using the synthetic myristoyl-d-Cys-tethered trimethoprim (mDcTMP) ligand. However, this system achieved PM-specific translocation only when the eDHFR tag was fused to the N terminus of proteins, thereby limiting its application. In this report, we engineered a universal PM-targeting tag for mDcTMP-induced protein translocation by grafting the hexalysine motif into an intra-loop region of eDHFR. We demonstrate the broad applicability of the new loop-engineered eDHFR tag and mDcTMP pair for conditional PM recruitment and activation of various tag-fused signaling proteins with different fusion configurations and for reversibly and repeatedly controlling protein localization to generate synthetic signal oscillations.
Volume 29(9)
Pages 1446-1464.e10
Published 2022-9-15
DOI 10.1016/j.chembiol.2022.06.005
PII S2451-9456(22)00237-9
PMID 35835118
MeSH Cell Membrane / metabolism Escherichia coli / metabolism Ligands Proteins Signal Transduction Tetrahydrofolate Dehydrogenase* / metabolism Trimethoprim* / pharmacology
IF 6.762
DNA material pCMV-VSV-G-RSV-Re (RDB04393)