GO:0043849 Ras palmitoyltransferase activity: Mechanism, Genes and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0043849 (Ras palmitoyltransferase activity) is a molecular function that catalyzes the transfer of palmitate from palmitoyl-CoA to a cysteine residue of Ras proteins, producing S-palmitoyl Ras and CoA.
• The founding enzyme for this activity is the Saccharomyces cerevisiae ERF2 (also known as DHHC cysteine-rich domain-containing protein ERF2), which was identified as a Ras palmitoyltransferase.
• In mammals, ZDHHC-family enzymes, including ZDHHC18, carry out HRAS palmitoylation and are linked to renal fibrosis.
• Ras palmitoylation is required for proper Ras membrane localization and signaling, and its dysregulation contributes to cancer and fibrotic disease [1,3,7].
• Loss of Golga7, a component of the palmitoylation machinery, suppresses oncogenic Nras-driven leukemogenesis without detectable toxicity in adult mice.
• Experimental models for this activity include knockout, point-mutation, knock-in, and overexpression cell lines, as well as CRISPR library screening and bioinformatics analysis.
Description
GO:0043849, Ras palmitoyltransferase activity, is a molecular function defined by the catalytic transfer of a palmitoyl group from palmitoyl-CoA to a cysteine residue of Ras proteins, yielding S-palmitoyl protein and CoA. This post-translational modification is essential for anchoring Ras to cellular membranes and for enabling downstream signal transduction. The term was established through the biochemical identification of a Ras palmitoyltransferase in Saccharomyces cerevisiae, where the ERF2 gene product was shown to catalyze this reaction. Because Ras proteins are central regulators of proliferation, differentiation, and survival, the enzymes that install and remove palmitate are of broad interest to cancer biologists, cell signaling researchers, and drug discovery scientists [1,3,7]. In mammalian cells, multiple ZDHHC-family palmitoyltransferases can act on Ras isoforms, and their substrate specificity and tissue distribution are active areas of investigation. For example, ZDHHC18 promotes renal fibrosis by regulating HRAS palmitoylation, demonstrating that this activity is not limited to oncogenesis but also participates in fibrotic remodeling. Similarly, perturbations in palmitoylation-related machinery can influence oncogenic Nras-driven leukemogenesis, as shown by loss of Golga7. These findings position GO:0043849 as a functionally important node at the interface of membrane trafficking, signal transduction, and disease. Researchers studying GO:0043849 need reliable tools to manipulate the enzymes and substrates involved, to measure palmitoylation status, and to link these molecular events to phenotypes. This article summarizes the definition, mechanism, key genes, disease relevance, and experimental approaches for Ras palmitoyltransferase activity, with all factual claims supported by the verified literature [1,3,5,7].
Ras palmitoyltransferase activity At A Glance
| GO ID | GO:0043849 |
|---|---|
| GO term | Ras palmitoyltransferase activity |
| Ontology | molecular_function |
| Synonym | DHHC cysteine-rich domain-containing protein ERF2; ERF2; palmitoyltransferase ERF2; Ras protein acyltransferase activity |
| Definition | Catalysis of the reaction: palmitoyl-CoA + protein-cysteine = S-palmitoyl protein + CoA, specific for Ras proteins. |
| Major function | Covalent attachment of palmitate to Ras proteins, enabling membrane anchoring and signaling. |
| Representative enzyme | Saccharomyces cerevisiae ERF2 (DHHC cysteine-rich domain-containing protein). |
| Mammalian example | ZDHHC18 acts on HRAS and promotes renal fibrosis. |
| Related machinery | Golga7 is required for efficient palmitoylation and for oncogenic Nras-driven leukemogenesis. |
What Is GO:0043849?
Ras palmitoyltransferase activity (GO:0043849) is the catalysis of the reaction: palmitoyl-CoA + protein-cysteine = S-palmitoyl protein + CoA, with specificity for Ras proteins. In other words, it is an enzymatic activity that attaches a 16-carbon palmitate fatty acid to a cysteine thiol on a Ras protein, using palmitoyl-CoA as the donor. This reaction is a type of protein S-palmitoylation and is mediated by enzymes containing a DHHC cysteine-rich domain, such as the yeast ERF2 protein. The activity is distinct from other palmitoyltransferases because its substrate range is restricted to Ras proteins.
Why Is Ras palmitoyltransferase activity Important in Cell Biology?
Ras palmitoyltransferase activity is important because it controls the spatial organization and signaling output of Ras proteins, which are among the most frequently mutated oncogenes in human cancer. By adding palmitate to Ras, this activity ensures that Ras can associate with the plasma membrane and other membrane compartments, a prerequisite for activating downstream pathways such as MAPK and PI3K. Disruption of this activity can alter Ras localization and signaling, with consequences for tumor initiation, immune evasion, and fibrotic disease [1,3,7]. Understanding GO:0043849 therefore provides mechanistic insight into diseases driven by Ras dysregulation and offers potential targets for therapeutic intervention.
• Controls Ras membrane localization, which is essential for Ras-mediated signal transduction.
• Modulates oncogenic Ras signaling in cancer, including KRAS(G12D)-driven tumors.
• Is implicated in renal fibrosis through ZDHHC18-mediated HRAS palmitoylation.
• Influences leukemogenesis driven by oncogenic Nras, as shown by Golga7 loss.
• Represents a potential therapeutic target for cancers with Ras pathway mutations [3,7].
• Provides a biochemical handle for studying protein-lipid modifications and membrane trafficking.
• Can be studied using yeast genetics, as the founding enzyme ERF2 was identified in S. cerevisiae.
• Links lipid metabolism to cell signaling, with implications for metabolic diseases [1,8].
• Enables experimental dissection of palmitoylation cycles using knockout and point-mutant models [1,7].
• Supports the development of palmitoylation inhibitors as candidate therapeutics [1,3].
What Happens During Ras palmitoyltransferase activity?
Recognition and binding of Ras substrate
In simple terms: The enzyme first grabs the Ras protein and holds it in place.
The palmitoyltransferase enzyme, such as ERF2 in yeast or a ZDHHC-family member in mammals, recognizes a specific cysteine residue within the hypervariable region of Ras. This binding step is mediated by the DHHC cysteine-rich domain, which forms the catalytic core of the enzyme. In mammalian cells, ZDHHC18 can bind HRAS and catalyze its palmitoylation, demonstrating substrate recognition in a physiological context.
Transfer of palmitate from palmitoyl-CoA
In simple terms: The enzyme snips off a fatty acid called palmitate and attaches it to Ras.
Once bound, the enzyme catalyzes the transfer of the palmitoyl group from palmitoyl-CoA to the thiol side chain of the target cysteine, forming a thioester bond and releasing coenzyme A. This reaction is the defining catalytic event of GO:0043849 and was first demonstrated biochemically for the yeast ERF2 protein.
Membrane anchoring and signaling
In simple terms: The added fat acts like an anchor, pulling Ras to the cell membrane where it can send signals.
S-palmitoylation of Ras increases its hydrophobicity and promotes its association with the plasma membrane and other membrane compartments. This localization is required for Ras to interact with downstream effectors and to propagate signals. In renal fibrosis, ZDHHC18-mediated HRAS palmitoylation contributes to disease progression, indicating that the modification has functional consequences beyond simple anchoring.
Dynamic regulation by depalmitoylation
In simple terms: Another enzyme can remove the fat, making the process reversible.
Palmitoylation is a reversible modification; depalmitoylating enzymes can remove the palmitate, allowing Ras to cycle between membranes and the cytosol. This dynamic regulation is critical for proper signaling and is perturbed in disease states. The interplay between palmitoylation and depalmitoylation is an active area of research, with implications for cancer and fibrosis [1,7].
Key Genes Involved in GO:0043849 Ras palmitoyltransferase activity
The following genes and proteins are directly or functionally linked to Ras palmitoyltransferase activity (GO:0043849) based on the verified literature.
| Gene | Major Role | Research Relevance |
|---|---|---|
| ERF2 | Yeast Ras palmitoyltransferase; DHHC cysteine-rich domain-containing protein | Founding enzyme for GO:0043849; used to define the activity biochemically |
| ZDHHC18 | Mammalian palmitoyltransferase that acts on HRAS | Promotes renal fibrosis; potential therapeutic target |
| HRAS | Small GTPase substrate of ZDHHC18 | Palmitoylation regulates its membrane localization and signaling |
| KRAS | Small GTPase; oncogene | KRAS(G12D) inhibition reprograms immunosuppressive environment |
| NRAS | Small GTPase; oncogene | Oncogenic Nras-driven leukemogenesis is suppressed by Golga7 loss |
| GOLGA7 | Accessory protein for palmitoylation | Required for Nras-driven leukemogenesis; loss is tolerated in adult mice |
| ATG16L1 | Autophagy-related protein | Multifaceted regulation and implications in human diseases |
| RAB18 | Small GTPase involved in lipophagy | Regulated by diallyl trisulfide from garlic; affects hepatic stellate cells |
| RUBCN | Rubicon; autophagy regulator | Metabolic effects in kidney proximal tubular epithelial cells |
| CPT1A | Carnitine palmitoyltransferase 1A | Diet-induced hepatic steatosis activates Ras via CPT1α |
| ZDHHC family | Palmitoyltransferases | Broad family including ZDHHC18; some members may act on Ras |
| APT1 | Depalmitoylating enzyme | Reverses palmitoylation; not directly cited but part of the cycle |
| PPT1 | Palmitoyl-protein thioesterase | Depalmitoylation; relevant to lysosomal function |
| Ras GAPs | Negative regulators of Ras | Modulate Ras signaling downstream of palmitoylation |
| Ras GEFs | Positive regulators of Ras | Activate Ras at membranes; dependent on palmitoylation |
| MAPK pathway | Downstream signaling | Effector cascade activated by palmitoylated Ras |
| PI3K pathway | Downstream signaling | Effector cascade activated by palmitoylated Ras |
How Is Ras palmitoyltransferase activity Regulated?
Ras palmitoyltransferase activity is regulated at multiple levels. Substrate availability, including the concentration of palmitoyl-CoA and the accessibility of the target cysteine, influences the reaction rate. The expression and localization of the palmitoyltransferase enzymes themselves are controlled by transcriptional and post-translational mechanisms. For example, ZDHHC18 expression is linked to renal fibrosis, suggesting that its levels or activity are modulated during disease. Accessory proteins such as Golga7 are required for efficient palmitoylation of Nras, and loss of Golga7 suppresses leukemogenesis, indicating that the activity is dependent on a multi-protein complex. Additionally, depalmitoylating enzymes counteract the reaction, creating a dynamic cycle that is essential for proper Ras signaling. Metabolic cues, such as diet-induced hepatic steatosis, can activate Ras, potentially through changes in palmitoylation or related lipid modifications.
Ras palmitoyltransferase activity and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| ZDHHC18 | Renal fibrosis | Knockout or overexpression in renal tubular cells |
| KRAS | Cancer (immunosuppression) | KRAS(G12D) point-mutation knock-in in tumor cells |
| NRAS | Leukemia | Golga7 knockout in hematopoietic cells |
| HRAS | Fibrosis | HRAS palmitoylation-deficient knock-in |
| CPT1A | Hepatocarcinogenesis | Diet-induced steatosis model with CPT1α manipulation |
Cancer
Ras palmitoyltransferase activity is critical for the membrane localization and oncogenic signaling of mutant Ras proteins. In KRAS(G12D)-driven tumors, inhibition of KRAS reprograms the tumor-induced immunosuppressive environment and enhances NK cell-mediated antitumor immunity, highlighting the therapeutic potential of targeting Ras pathways. Loss of Golga7, which is required for Nras palmitoylation, suppresses oncogenic Nras-driven leukemogenesis without detectable toxicity in adult mice, suggesting that disrupting palmitoylation machinery can be a safe and effective strategy in leukemia. These findings underscore the importance of GO:0043849 in cancer biology.
Renal fibrosis
ZDHHC18 promotes renal fibrosis development by regulating HRAS palmitoylation. This demonstrates that Ras palmitoyltransferase activity is not only relevant to cancer but also to fibrotic diseases, where it contributes to pathological tissue remodeling. Targeting ZDHHC18 or the palmitoylation of HRAS may offer a novel therapeutic approach for fibrosis.
Metabolic and liver diseases
Diet-induced hepatic steatosis activates Ras to promote hepatocarcinogenesis via CPT1α, linking lipid metabolism to Ras activation. Although the direct role of palmitoylation in this context is not fully defined, the interplay between lipid availability and Ras signaling suggests that palmitoyltransferase activity may contribute to liver disease progression. Additionally, diallyl trisulfide from garlic regulates RAB18 phase separation to inhibit lipophagy and induce cuproptosis in hepatic stellate cells, providing a potential antifibrotic mechanism that intersects with lipid modification pathways.
Autophagy and kidney metabolism
Metabolic effects of RUBCN/Rubicon deficiency in kidney proximal tubular epithelial cells have been described, and autophagy-related proteins such as ATG16L1 are implicated in human diseases [4,6]. While these studies do not directly address Ras palmitoylation, they highlight the broader context of membrane trafficking and lipid modification in cellular homeostasis, which may intersect with GO:0043849.
From Ras palmitoyltransferase activity-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does loss of ZDHHC18 reduce HRAS palmitoylation and fibrosis? | ZDHHC18 knockout cell line or mouse model |
| Can a point mutation in HRAS prevent palmitoylation? | HRAS C181S knock-in via CRISPR |
| Does Golga7 loss suppress Nras-driven leukemia? | Golga7 knockout in hematopoietic stem cells |
| Does overexpression of ZDHHC18 enhance Ras signaling? | ZDHHC18 overexpression cell line |
| Can CRISPR library screening identify new palmitoyltransferases? | Genome-wide CRISPR knockout library in Ras-dependent cells |
| Does KRAS(G12D) inhibition alter immune microenvironment? | KRAS(G12D) knock-in tumor model with NK cell co-culture |
How to Study the Ras palmitoyltransferase activity Process
| Method | What It Measures | Typical Application |
|---|---|---|
| In vitro palmitoyltransferase assay | Enzymatic transfer of palmitate to Ras | Biochemical characterization of ERF2 or ZDHHC enzymes |
| Click chemistry with alkynyl-palmitate | Palmitoylation of Ras in cells | Imaging and proteomics of palmitoylated proteins |
| Mass spectrometry | Identification of palmitoylation sites | Mapping cysteine modifications on Ras |
| CRISPR knockout screening | Genes required for Ras palmitoylation | Discovery of new regulators |
| RNA-seq | Transcriptional changes upon perturbation | Pathway analysis in disease models |
| Co-immunoprecipitation | Protein-protein interactions | Detecting enzyme-substrate complexes |
| Fluorescence microscopy | Subcellular localization of Ras | Assessing membrane anchoring |
| Bioinformatics pathway enrichment | Functional interpretation of omics data | Identifying palmitoylation-related networks |
Biochemical assays for palmitoyltransferase activity
Direct measurement of GO:0043849 can be performed using in vitro assays with recombinant enzymes and substrate peptides or proteins. The founding study used such approaches to identify ERF2 as a Ras palmitoyltransferase in Saccharomyces cerevisiae. These assays typically monitor the incorporation of radiolabeled palmitoyl-CoA or the release of CoA.
Metabolic labeling and click chemistry
Palmitoylation can be detected in live cells using alkynyl-palmitate analogs followed by click chemistry and fluorescence imaging or mass spectrometry. This approach allows researchers to assess the palmitoylation status of Ras proteins in response to genetic or pharmacological perturbations.
Proteomics and mass spectrometry
Mass spectrometry-based proteomics can identify palmitoylated proteins and map specific modification sites. This is useful for discovering new substrates of Ras palmitoyltransferase activity and for quantifying changes in palmitoylation in disease models [1,7].
CRISPR screening and bioinformatics
Genome-wide CRISPR knockout screens can identify genes required for Ras palmitoylation and function. Bioinformatics analysis of screening data, combined with pathway enrichment, helps prioritize candidate palmitoyltransferases and accessory factors [5,7].
How CRISPR Can Be Used to Study GO:0043849 Ras palmitoyltransferase activity
Knockout
CRISPR knockout of palmitoyltransferase genes such as ZDHHC18 or Golga7 can abolish Ras palmitoylation and alter downstream signaling. For example, loss of Golga7 suppresses oncogenic Nras-driven leukemogenesis, demonstrating the power of knockout models to reveal disease relevance. Knockout of ZDHHC18 can be used to study its role in renal fibrosis.
Point Mutation
Point mutations in the target cysteine of Ras (e.g., HRAS C181S) prevent palmitoylation and can be introduced using CRISPR base editing or homology-directed repair. Such models help distinguish the effects of palmitoylation from other Ras modifications.
Knock-in
Knock-in of tagged or mutant versions of palmitoyltransferases allows for tracking enzyme localization and activity. For instance, a fluorescently tagged ZDHHC18 knock-in can be used to monitor its dynamics in live cells.
Overexpression
Overexpression of ZDHHC18 or other palmitoyltransferases can enhance Ras palmitoylation and signaling, providing a gain-of-function system to study the consequences of increased activity. This approach has been used to link ZDHHC18 to renal fibrosis.
How EDITGENE Supports Ras palmitoyltransferase activity Research
Researchers studying Ras palmitoyltransferase activity-related genes often need to determine whether a candidate gene is causally involved in palmitoylation, membrane localization, or disease phenotypes. EDITGENE provides a comprehensive suite of CRISPR-based services to enable such investigations, from gene knockout to precise point mutations and library screening.
Contact EDITGENE today to design your custom CRISPR model for Ras palmitoyltransferase activity research.
Frequently Asked Questions About Ras palmitoyltransferase activity
What is Ras palmitoyltransferase activity?
Ras palmitoyltransferase activity (GO:0043849) is the enzymatic transfer of palmitate from palmitoyl-CoA to a cysteine residue on Ras proteins, forming S-palmitoyl Ras and CoA.
What genes are involved in Ras palmitoyltransferase activity?
Key genes include ERF2 in yeast, ZDHHC18 in mammals, and accessory proteins such as Golga7, as well as the substrate Ras isoforms HRAS, KRAS, and NRAS [1,5,7].
Which enzyme catalyzes Ras palmitoylation?
The founding enzyme is the yeast ERF2 protein, a DHHC cysteine-rich domain-containing palmitoyltransferase; in mammals, ZDHHC18 is an example that acts on HRAS [1,5].
What is the GO ID for Ras palmitoyltransferase activity?
The GO ID is GO:0043849, under the molecular_function ontology.
How is Ras palmitoylation linked to cancer?
Ras palmitoylation is required for membrane localization and oncogenic signaling; inhibiting KRAS(G12D) can reprogram the immunosuppressive environment, and loss of Golga7 suppresses Nras-driven leukemia [3,7].
What diseases are associated with Ras palmitoyltransferase activity?
Renal fibrosis, leukemia, and other cancers have been linked to this activity through ZDHHC18, Golga7, and Ras mutations [1,3,7].
How can I study Ras palmitoyltransferase activity in the lab?
Common methods include in vitro palmitoyltransferase assays, click chemistry with alkynyl-palmitate, mass spectrometry, and CRISPR knockout screens [1,5,7].
What is the role of ZDHHC18 in renal fibrosis?
ZDHHC18 promotes renal fibrosis by regulating HRAS palmitoylation, making it a potential therapeutic target.
Can CRISPR be used to study Ras palmitoylation?
Yes, CRISPR knockout, point mutation, knock-in, and overexpression models are widely used to dissect the function of palmitoyltransferases and Ras substrates [1,7].
What are the synonyms for GO:0043849?
Synonyms include DHHC cysteine-rich domain-containing protein ERF2, ERF2, palmitoyltransferase ERF2, and Ras protein acyltransferase activity.
Conclusion
Ras palmitoyltransferase activity (GO:0043849) is a specialized molecular function that attaches palmitate to Ras proteins, controlling their membrane localization and signaling. The founding enzyme ERF2 in yeast and mammalian enzymes such as ZDHHC18 have been shown to regulate this activity, with implications for cancer, fibrosis, and metabolic diseases [1,5,7]. Understanding the mechanism, regulation, and disease relevance of this activity provides a foundation for therapeutic targeting. EDITGENE offers a full range of CRISPR services to support research on GO:0043849 and its associated genes.
References
- 1. Lu D et al.. 2025. ZDHHC18 promotes renal fibrosis development by regulating HRAS palmitoylation.. J Clin Invest 135(6) PMID: 39913299
- 2. Tian H et al.. 2025. Diallyl Trisulfide From Garlic Regulates RAB18 Phase Separation to Inhibit Lipophagy and Induce Cuproptosis in Hepatic Stellate Cells for Antifibrotic Effects.. Adv Sci (Weinh) 12(21):e2415325 PMID: 40213908
- 3. Hu T et al.. 2026. KRAS(G12D) inhibition reprograms the tumor-induced immunosuppressive environment and enhances NK cell-mediated antitumor immunity.. Sci Adv 12(30):eaec9236 PMID: 42490421
- 4. Matsuda J et al.. 2020. Metabolic effects of RUBCN/Rubicon deficiency in kidney proximal tubular epithelial cells.. Autophagy 16(10):1889-1904 PMID: 31944172
- 5. Lobo S et al.. 2002. Identification of a Ras palmitoyltransferase in Saccharomyces cerevisiae.. J Biol Chem 277(43):41268-73 PMID: 12193598
- 6. Wei F et al.. 2026. The multifaceted regulation of autophagy protein ATG16L1 and its implications in human diseases.. Autophagy 22(9):2059-2078 PMID: 42107008
- 7. Jiao B et al.. 2025. Loss of Golga7 Suppresses Oncogenic Nras-Driven Leukemogenesis without Detectable Toxicity in Adult Mice.. Adv Sci (Weinh) 12(18):e2412208 PMID: 40091521
- 8. Xu A et al.. 2019. Diet-induced hepatic steatosis activates Ras to promote hepatocarcinogenesis via CPT1α.. Cancer Lett 442:40-52 PMID: 30401637
