| 2016 |
DHHC20 palmitoylates EGFR at cysteine residues within its unstructured C-terminal tail; this palmitoylation 'pins' the C-terminal tail to the plasma membrane, impeding EGFR activation. Mutation of these cysteine residues to alanine is sufficient to activate EGFR signaling, promote cell migration and transformation, and sensitize cells to EGFR tyrosine kinase inhibition. |
Mass spectrometry identification of palmitoylated cysteines, site-directed mutagenesis (Cys→Ala), loss-of-function (DHHC20 inhibition), cell migration and transformation assays |
Molecular cell |
High |
27153536
|
| 2017 |
ZDHHC20 enhances IFITM3 palmitoylation and uniquely increases IFITM3 antiviral activity (against influenza) when both are co-overexpressed. ZDHHC20 co-localizes with IFITM3 at lysosomes, unlike ZDHHC3/7/15 which show perinuclear localization, suggesting that the subcellular site of palmitoylation influences IFITM3 activity. Combined knockdown of ZDHHC3 and ZDHHC7 in ZDHHC20-knockout cells was required to reduce endogenous IFITM3 palmitoylation, demonstrating functional redundancy among ZDHHCs. |
Overexpression screen (23 mammalian ZDHHCs), ZDHHC knockout cell line library, siRNA knockdown, metabolic palmitoylation labeling, viral infection assay, co-localization by fluorescence microscopy |
The Journal of biological chemistry |
High |
29079573
|
| 2019 |
Human DHHC20 is catalytically active when reconstituted in POPC nanodiscs. Molecular dynamics simulations reveal that DHHC20 induces a drastic deformation/invagination of the cytoplasmic leaflet of the lipid membrane, causing the catalytic Cys (within the DHHC motif) to become hydrated and optimally positioned to undergo autoacylation upon encountering acyl-CoA. This membrane reshaping resolves the paradox of a nucleophilic Cys being exposed to a hydrophobic membrane environment. |
Biochemical reconstitution in POPC nanodiscs, microsecond all-atom molecular dynamics simulations |
Biophysical journal |
Medium |
31858978
|
| 2021 |
ZDHHC20 mediates S-acylation of the ORAI1 Ca2+ channel at Cys143. This palmitoylation targets ORAI1 to cholesterol-rich lipid raft domains and is required for efficient TCR recruitment and signaling at the immune synapse. Cys143 mutations reduced ORAI1 currents, store-operated Ca2+ entry, NFATC1 translocation, and IL-2 secretion evoked by TCR engagement; the acylation-deficient channel was also recruited less efficiently to the immune synapse along with actin and TCR. |
Site-directed mutagenesis (Cys143), patch-clamp electrophysiology, Ca2+ imaging, NFATC1 nuclear translocation assay, IL-2 ELISA, enforced ZDHHC20 overexpression, lipid raft fractionation, live-cell fluorescence microscopy at immune synapse |
eLife |
High |
34913437
|
| 2022 |
Molecular dynamics of membrane-embedded hDHHC20 shows that among acyl-CoA chains of various lengths, only C16 (palmitoyl) adopts a conformation suitable for hDHHC20 autoacylation within the hydrophobic cavity formed by four transmembrane helices. A V185G mutant shifts preference to C18, demonstrating that residues at the cavity ceiling determine acyl-chain length selectivity. An unusual hydrophilic ridge in TM helix 4 of DHHC20 may mediate association with substrate protein TM domains. |
Molecular dynamics simulation of membrane-embedded hDHHC20 and mutants with various acyl-CoAs; mutagenesis of cavity residues; experimental validation of spike protein acylation by S→A mutagenesis |
International journal of molecular sciences |
Medium |
35563480
|
| 2024 |
ZDHHC20 palmitoylates YTHDF3 at Cys474, preventing its chaperone-mediated autophagic (CMA) degradation and causing accumulation of YTHDF3 protein. Stabilized YTHDF3 in turn leads to abnormal accumulation of MYC mRNA/protein, promoting malignant phenotypes in pancreatic cancer. A YTHDF3-derived peptide competitively inhibiting ZDHHC20-mediated YTHDF3 palmitoylation downregulates MYC and inhibits KRAS-mutant pancreatic cancer progression. |
KPC mouse model, acyl-biotin exchange assay, site-directed mutagenesis (Cys474), co-immunoprecipitation, protein half-life assay, competitive peptide inhibitor, in vivo tumor models |
Nature communications |
High |
38821916
|
| 2024 |
ZDHHC20 palmitoylates fatty acid synthase (FASN) at Cys1471 and Cys1881. This palmitoylation competes with ubiquitination at these sites, blocking FASN degradation via the E3 ubiquitin ligase complex SNX8-TRIM28. ZDHHC20 knockout or pharmacological inhibition, or FASN C1471S/C1881S mutation, accelerates FASN degradation and reduces hepatocarcinogenesis in two chemical carcinogen mouse models. |
ZDHHC20 knockout mice (DEN and DEN/CCl4 HCC models), palmitoylation LC-MS, acyl-biotin exchange assay, co-immunoprecipitation, ubiquitination assay, protein half-life assay, site-directed mutagenesis (C1471S/C1881S), immunofluorescence microscopy |
Molecular cancer |
High |
39696259
|
| 2024 |
zDHHC20 palmitoylates CD80 at Cys261/262/266/271 (transmembrane and cytoplasmic regions). This palmitoylation protects CD80 from ubiquitin-mediated proteasomal degradation, ensures accurate plasma membrane localization, and is required for CD80 costimulatory function in T cell activation. Palmitoylation-deficient CD80 (4CS mutant) loses membrane localization and fails to costimulate T cells. |
Metabolic palmitoylation labeling, site-directed mutagenesis (4CS), co-immunoprecipitation, ubiquitination assay, protein stability assay, immunofluorescence, T cell co-stimulation assay |
Acta pharmacologica Sinica |
High |
38467718
|
| 2024 |
ZDHHC20 was identified in an in vivo shRNA screen as critical for pancreatic cancer metastatic outgrowth. This pro-metastatic function is abrogated in immunocompromised animals and in animals depleted of NK cells, placing ZDHHC20-dependent palmitoylation in the regulation of tumor-innate immune interactions required for distant metastasis. A chemical genetics substrate profiling platform identified multiple novel ZDHHC20 substrates. |
In vivo shRNA screen, immunocompromised and NK-cell-depleted mouse models, chemical genetics substrate profiling platform |
Cell reports |
Medium |
38733589
|
| 2024 |
ZDHHC20 overexpression increases phosphorylation of PI3K and AKT in hepatocellular carcinoma cells, while ZDHHC20 knockdown decreases PI3K and AKT phosphorylation. PI3K-AKT pathway inhibitors (LY294002 and MK2206) block the pro-proliferative effect of ZDHHC20 overexpression, placing ZDHHC20 upstream of the PI3K-AKT pathway in HCC. |
siRNA knockdown, overexpression, Western blotting for PI3K/AKT phosphorylation, pharmacological inhibition, xenograft mouse model |
Journal of hepatocellular carcinoma |
Low |
39309302
|
| 2025 |
ZDHHC20 palmitoylates KAP1/TRIM28 at Cys232. This palmitoylation increases chromatin binding of phosphorylated KAP1, enhancing chromatin accessibility and recruitment of DNA damage response components BRCA1 and 53BP1. ATM kinase phosphorylates ZDHHC20 at Ser339 in response to DNA damage, which in turn increases KAP1 palmitoylation, defining an ATM-ZDHHC20-KAP1 axis in the DNA damage response. ZDHHC20 knockout enhances radiosensitivity in mice, tumor cell lines, and xenograft models. |
ZDHHC family screen, palmitoylation label-free quantitative proteomics, acyl-biotin exchange (ABE) assay, site-directed mutagenesis (Cys232), chromatin fractionation, BRCA1/53BP1 recruitment assay, ATM phosphorylation site mapping (Ser339), ZDHHC20 knockout mice, xenograft models, radiosensitivity assays |
Oncogene |
High |
41109928
|
| 2026 |
MEF2A transcriptionally activates ZDHHC20 (validated by dual-luciferase reporter and ChIP assays). ZDHHC20 in turn activates the NF-κB signaling pathway, promoting AML cell proliferation and doxorubicin resistance. Overexpression of ZDHHC20 reverses the antitumor effects of MEF2A silencing. |
Dual-luciferase reporter assay, chromatin immunoprecipitation (ChIP), siRNA knockdown, overexpression rescue, Western blotting, xenograft mouse model |
Biology direct |
Medium |
41882733
|
| 2026 |
ZDHHC20 palmitoylates CMPK2 at Cys137 and Cys153, maintaining CMPK2 mitochondrial localization. This palmitoylation is reversed by the thioesterase PPT1. CMPK2 palmitoylation supports ddhCTP production and MAVS stabilization, enabling IFN-I production and antiviral immunity. Palmitic acid activates this pathway; PPT1 deficiency restores CMPK2 palmitoylation and antiviral immunity. |
Chemical library screen, palmitoylation assay, site-directed mutagenesis (Cys137/153), subcellular fractionation/mitochondrial localization assay, CMPK2 knockout, PPT1 knockout/inhibition, IFN-I production assay, viral replication assay, in vivo diet and pharmacological intervention |
Advanced science |
High |
42011944
|
| 2026 |
RAB11A acts as a scaffold that recruits ZDHHC20 to promote FGFR3 palmitoylation, inhibiting FGFR3 degradation and retaining it at the plasma membrane in bladder cancer cells. SREBP2 directly activates RAB11A transcription (dependent on its liquid-liquid phase separation capability), placing ZDHHC20 downstream of a SREBP2-RAB11A axis. Knockdown of ZDHHC20 or RAB11A reduced membrane FGFR3 and attenuated tumor growth in xenografts. |
Co-immunoprecipitation (RAB11A-ZDHHC20 interaction), promoter-reporter assay (SREBP2→RAB11A), SREBP2 phase-separation mutant (F178A), RAB11A/ZDHHC20 knockdown, immunoblotting for membrane FGFR3, xenograft tumor models |
Cellular oncology |
Medium |
41586981
|
| 2024 |
A selective, orally bioavailable small-molecule inhibitor SD-066-4 inhibits ZDHHC20 acyltransferase activity. A specific alanine residue in ZDHHC20 accommodates the methyl group of SD-066-4, providing isoform selectivity over other ZDHHCs. SD-066-4 stably reduces EGFR S-acylation in KRAS-mutant cells and blocks growth of KRAS-mutant lung tumors, extending overall survival in a mouse model. |
Selective inhibitor identification, mutagenesis identifying selectivity-determining alanine residue, EGFR palmitoylation assay, KRAS-mutant lung tumor mouse model, survival analysis |
bioRxivpreprint |
Medium |
bio_10.1101_2024.07.18.604152
|