GO:0035331 negative regulation of hippo signaling: Pathway, Genes, Functions and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0035331 (negative regulation of hippo signaling) describes any process that stops, prevents, or reduces the frequency, rate or extent of hippo signaling.
• The Hippo pathway is a kinase cascade in which upstream signals control YAP/TAZ phosphorylation, localization and transcriptional output.
• Negative regulation of Hippo signaling therefore increases YAP/TAZ nuclear activity and promotes target gene expression.
• Dysregulated negative regulation of Hippo signaling is implicated in breast cancer, bladder cancer, esophageal squamous carcinoma and musculoskeletal disorders [1,3,5,6].
• E3 ubiquitin ligases such as PARK2 and RNF187 can modulate Hippo signaling and tumor progression, illustrating crosstalk with ubiquitination [6,7].
• CRISPR knockout, point mutation, knock-in and overexpression models are essential to test causality of candidate negative regulators of Hippo signaling [1,2,8].
Description
GO:0035331, negative regulation of hippo signaling, is a biological process term that captures any mechanism which stops, prevents, or reduces the frequency, rate or extent of hippo signaling. The Hippo pathway is an evolutionarily conserved kinase cascade that controls cell proliferation, apoptosis and organ size by regulating the transcriptional co-activators YAP and TAZ. When Hippo signaling is active, YAP/TAZ are phosphorylated and retained in the cytoplasm; when Hippo signaling is negatively regulated, YAP/TAZ translocate to the nucleus and drive target gene expression. This term is therefore central to understanding how cells bypass growth suppression. Researchers study GO:0035331 because its dysregulation is linked to multiple human diseases, including breast cancer, bladder cancer, esophageal squamous carcinoma and musculoskeletal disorders [1,3,5,6]. In breast cancer, reciprocal regulation between microRNAs and Hippo-YAP/TAZ signaling influences precision medicine approaches. In bladder cancer, circXRN2 suppresses tumor progression by activating the Hippo pathway, highlighting how negative regulators of Hippo signaling can be therapeutically relevant. In esophageal squamous carcinoma, the E3 ubiquitin ligase PARK2 regulates Hippo/YAP signaling and disease progression. Understanding the molecular players that negatively regulate Hippo signaling is thus essential for both basic developmental biology and translational oncology.
negative regulation of hippo signaling At A Glance
| GO ID | GO:0035331 |
|---|---|
| GO term | negative regulation of hippo signaling |
| Ontology | biological_process |
| Synonym | negative regulation of hippo signaling cascade; negative regulation of hippo signaling pathway; negative regulation of hippo signalling cascade |
| Major function | Reduces or prevents Hippo signaling, thereby promoting YAP/TAZ transcriptional activity |
| Related process | Hippo signaling pathway, YAP/TAZ regulation, organ size control |
| Disease relevance | Breast cancer, bladder cancer, esophageal squamous carcinoma, musculoskeletal disorders [1,3,5,6] |
| Key regulators | E3 ubiquitin ligases such as PARK2 and RNF187, circular RNAs, microRNAs [3,5,6,7] |
| Research models | CRISPR knockout, point mutation, knock-in, overexpression, library screening [1,2,8] |
What Is GO:0035331?
In our own words, GO:0035331 refers to any cellular process that reduces, blocks or prevents the activity of the Hippo signaling cascade. Because Hippo signaling normally restrains the oncogenic co-activators YAP and TAZ, negative regulation of this pathway typically results in increased YAP/TAZ nuclear accumulation and enhanced transcription of growth-promoting genes. This regulation can occur through direct inhibition of Hippo kinases, modulation of upstream polarity or junctional cues, or crosstalk with other signaling pathways.
Why Is negative regulation of hippo signaling Important in Cell Biology?
Negative regulation of Hippo signaling is important because it determines whether cells interpret upstream growth-suppressive cues as active or silenced. When this process is aberrantly activated, YAP/TAZ become constitutively nuclear and drive proliferation, survival and migration, contributing to tumorigenesis and other pathologies [1,3,5,6]. Conversely, understanding how to enhance negative regulation of Hippo signaling or restore Hippo activity is a therapeutic goal in cancers where YAP/TAZ are hyperactive [3,4,5]. The term also matters for regenerative medicine and musculoskeletal biology, where controlled YAP/TAZ activity influences tissue repair and osteopotential [1,2].
• Controls YAP/TAZ nuclear localization and transcriptional output.
• Influences cell proliferation, apoptosis and organ size.
• Implicated in breast cancer progression and precision medicine.
• Linked to bladder cancer tumor progression through circXRN2 and histone lactylation.
• Regulates esophageal squamous carcinoma progression via PARK2.
• Modulates triple-negative breast cancer progression through RNF187.
• Affects musculoskeletal disorders and bone marrow cell osteopotential [1,2].
• Provides targets for CRISPR-based functional genomics and drug discovery [1,8].
What Happens During negative regulation of hippo signaling?
Upstream signal integration and kinase cascade
In simple terms: The Hippo pathway is a relay of kinases that normally puts brakes on growth; negative regulation removes or weakens those brakes.
Hippo signaling is a kinase cascade in which upstream signals converge on MST1/2 and LATS1/2 kinases, which phosphorylate YAP/TAZ and promote their cytoplasmic retention or degradation. Negative regulation of Hippo signaling can occur when upstream cues such as cell density, polarity or mechanical forces are altered, leading to reduced kinase activity and subsequent YAP/TAZ dephosphorylation.
YAP/TAZ dephosphorylation and nuclear translocation
In simple terms: When the brakes are off, YAP/TAZ move into the nucleus to turn on growth genes.
Reduced Hippo kinase activity leads to YAP/TAZ dephosphorylation, allowing them to translocate to the nucleus and partner with TEAD transcription factors to activate target genes. This step is a key functional consequence of negative regulation of Hippo signaling and is often measured as a readout of pathway activity.
Multiphase coalescence and signalosome assembly
In simple terms: Cells can switch the pathway on or off by clustering proteins into droplets, a process called phase separation.
Multiphase coalescence mediates Hippo pathway activation, demonstrating that dynamic assembly of signaling condensates can regulate pathway output. Negative regulation of Hippo signaling may involve disruption or remodeling of these condensates, thereby reducing kinase activation.
Crosstalk with ubiquitination and degradation
In simple terms: Tagging proteins with ubiquitin can remove them and shut down the pathway.
E3 ubiquitin ligases such as PARK2 and RNF187 regulate Hippo/YAP signaling and tumor progression, indicating that ubiquitin-mediated degradation of pathway components is a mechanism of negative regulation [6,7]. For example, PARK2 regulates Hippo/YAP signaling in esophageal squamous carcinoma, and RNF187 regulates Hippo signaling in triple-negative breast cancer [6,7].
Non-coding RNA and epigenetic modulation
In simple terms: Small RNAs and chemical marks on DNA or proteins can also turn the pathway down.
MicroRNAs reciprocally regulate Hippo-YAP/TAZ signaling in breast cancer, and circular RNA circXRN2 suppresses tumor progression by activating the Hippo pathway in bladder cancer [3,5]. Histone lactylation is involved in circXRN2-mediated Hippo pathway activation, illustrating epigenetic control of negative regulation.
Key Genes Involved in GO:0035331 negative regulation of hippo signaling
The following genes and proteins are experimentally implicated in negative regulation of Hippo signaling or in the Hippo pathway itself, based on the verified literature.
| Gene | Major Role | Research Relevance |
|---|---|---|
| YAP1 | Transcriptional co-activator downstream of Hippo signaling | Readout of negative regulation; nuclear localization indicates pathway inhibition |
| WWTR1 (TAZ) | Transcriptional co-activator downstream of Hippo signaling | Readout of negative regulation; nuclear localization indicates pathway inhibition |
| STK3/STK4 (MST1/2) | Core Hippo kinases | Phosphorylate LATS1/2; their inhibition constitutes negative regulation |
| LATS1 | Core Hippo kinase | Phosphorylates YAP/TAZ; reduced activity indicates negative regulation |
| LATS2 | Core Hippo kinase | Phosphorylates YAP/TAZ; reduced activity indicates negative regulation |
| PARK2 | E3 ubiquitin ligase | Regulates Hippo/YAP signaling and esophageal squamous carcinoma progression |
| RNF187 | E3 ubiquitin ligase | Regulates Hippo signaling and triple-negative breast cancer progression |
| CircXRN2 | Circular RNA | Suppresses tumor progression by activating the Hippo pathway in bladder cancer |
| MIRNAs (e.g., miR-), microRNAs | Post-transcriptional regulators | Reciprocal regulation with Hippo-YAP/TAZ in breast cancer |
| TEAD1-4 | Transcription factors partnering YAP/TAZ | Mediate transcriptional output upon negative regulation |
| NF2 | Upstream Hippo pathway activator | Loss of NF2 can lead to negative regulation of Hippo signaling |
| WWC1 (KIBRA) | Upstream Hippo pathway regulator | Modulates Hippo kinase activity |
| AMOT | Junctional polarity protein | Influences Hippo signaling via upstream cues |
| RASSF1 | Ras association domain family member | Can modulate Hippo signaling |
| SAV1 | Scaffold protein for MST kinases | Required for Hippo kinase activation |
| MOB1A/B | LATS kinase co-activators | Required for Hippo signaling; loss promotes negative regulation |
| Rauvolfia tetraphylla compounds | Plant-derived apoptosis regulators | Regulate Hippo signaling-mediated apoptosis in triple-negative breast cancer |
How Is negative regulation of hippo signaling Regulated?
Negative regulation of Hippo signaling is itself regulated by multiple inputs, including cell density, mechanical forces, polarity cues, ubiquitination, non-coding RNAs and epigenetic modifications [3,5,6,7,8]. For example, E3 ubiquitin ligases PARK2 and RNF187 modulate Hippo signaling and tumor progression, while microRNAs and circular RNAs provide post-transcriptional control [3,5,6,7]. Multiphase coalescence of signaling components can also switch the pathway on or off, adding a biophysical layer of regulation.
negative regulation of hippo signaling and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| RNF187 | Triple-negative breast cancer progression | CRISPR knockout in TNBC cell lines |
| PARK2 | Esophageal squamous carcinoma progression | Knockout or overexpression in ESCC cells |
| CircXRN2 | Bladder cancer tumor progression | Overexpression or knockdown in bladder cancer cells |
| YAP1/WWTR1 | Musculoskeletal disorders and osteopotential | Knockout or knock-in in bone marrow cells [1,2] |
| MicroRNAs | Breast cancer precision medicine | Mimic or inhibitor transfection in breast cancer cells |
Breast cancer
Hippo-YAP/TAZ signaling is reciprocally regulated by microRNAs in breast cancer, and this crosstalk has implications for precision medicine. RNF187 regulates Hippo signaling and triple-negative breast cancer progression, and Rauvolfia tetraphylla compounds regulate Hippo signaling-mediated apoptosis in triple-negative breast cancer [4,7]. Negative regulation of Hippo signaling therefore contributes to breast cancer growth and survival [3,4,7].
Bladder cancer
CircXRN2 suppresses tumor progression driven by histone lactylation through activating the Hippo pathway in human bladder cancer. This indicates that negative regulation of Hippo signaling, or loss of Hippo activation, promotes bladder cancer progression.
Esophageal squamous carcinoma
The E3 ubiquitin ligase PARK2 regulates Hippo/YAP signaling and esophageal squamous carcinoma progression. Dysregulation of this axis represents a mechanism by which negative regulation of Hippo signaling may contribute to esophageal cancer.
Musculoskeletal disorders
The Hippo-YAP/TAZ signaling pathway plays emerging roles in musculoskeletal disorders, and Hippo signaling is involved in osteopotential regulation of murine bone marrow cells under simulated microgravity [1,2]. Negative regulation of Hippo signaling may therefore influence bone and joint biology [1,2].
From negative regulation of hippo signaling-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Is a candidate gene a negative regulator of Hippo signaling? | CRISPR knockout cell line followed by YAP/TAZ localization assay |
| Does a specific point mutation alter Hippo pathway regulation? | Point-mutation knock-in via CRISPR |
| Does a candidate gene product interact with Hippo kinases? | Tagged knock-in for co-immunoprecipitation |
| Does overexpression of a gene reduce Hippo signaling? | CRISPR overexpression or lentiviral overexpression |
| Which genes regulate Hippo signaling in a disease context? | CRISPR library screening in cancer cell lines [1,8] |
| Does a non-coding RNA modulate Hippo signaling? | Overexpression or knockout of circRNA/miRNA [3,5] |
How to Study the negative regulation of hippo signaling Process
| Method | What It Measures | Typical Application |
|---|---|---|
| CRISPR knockout | Loss-of-function effects on Hippo signaling | Identify negative regulators |
| Point mutation knock-in | Effect of specific amino acid changes | Test functional domains |
| RNA-seq | Transcriptional changes in YAP/TAZ targets | Pathway output profiling |
| Proteomics | Protein interactions and modifications | Identify pathway components |
| Immunofluorescence | YAP/TAZ nuclear vs cytoplasmic localization | Readout of pathway activity |
| CRISPR library screening | Pooled gene function in pathway regulation | Discover novel regulators [1,8] |
| Bioinformatics | Pathway enrichment and network analysis | Prioritize candidate genes |
CRISPR knockout and point mutation
CRISPR knockout of candidate genes followed by YAP/TAZ localization and target gene expression analysis can determine whether a gene negatively regulates Hippo signaling. Point mutations can be introduced to test specific residues required for pathway modulation.
Transcriptional and proteomic profiling
RNA-seq and proteomics can identify global changes in YAP/TAZ target genes and interacting proteins upon perturbation of negative regulators. This is useful for mapping downstream effects of Hippo pathway inhibition.
Imaging and localization assays
Immunofluorescence for YAP/TAZ nuclear localization is a standard readout of Hippo pathway activity and negative regulation. Live-cell imaging of signaling condensates can reveal dynamic regulation.
Functional screens and bioinformatics
CRISPR library screening combined with bioinformatics can identify novel negative regulators of Hippo signaling in a disease-relevant context [1,8]. Pathway enrichment and network analysis help prioritize candidates.
How CRISPR Can Be Used to Study GO:0035331 negative regulation of hippo signaling
Knockout
CRISPR knockout of a candidate gene can test whether it is required for negative regulation of Hippo signaling; loss of a negative regulator should reduce YAP/TAZ nuclear localization.
Point Mutation
Point mutation knock-in can dissect which residues of a protein are essential for modulating Hippo signaling, for example by disrupting a phosphorylation site or interaction interface.
Knock-in
Tagged knock-in allows endogenous labeling of Hippo pathway components for imaging, co-immunoprecipitation and dynamic studies of negative regulation.
Overexpression
CRISPR overexpression or lentiviral overexpression of a candidate gene can test whether increased dosage is sufficient to reduce Hippo signaling and activate YAP/TAZ.
How EDITGENE Supports negative regulation of hippo signaling Research
Researchers studying negative regulation of hippo signaling-related genes often need to determine whether a candidate gene is causally involved in pathway modulation or simply correlated with YAP/TAZ activity. EDITGENE provides the CRISPR tools and services to move from correlation to causation.
Contact EDITGENE today to design your custom CRISPR model for negative regulation of hippo signaling research.
Frequently Asked Questions About negative regulation of hippo signaling
What is negative regulation of hippo signaling?
It is any process that stops, prevents, or reduces the frequency, rate or extent of hippo signaling, often leading to increased YAP/TAZ activity.
What is GO:0035331?
GO:0035331 is the Gene Ontology identifier for negative regulation of hippo signaling, a biological process term.
What genes are involved in negative regulation of hippo signaling?
Genes include YAP1, WWTR1 (TAZ), STK3/STK4, LATS1/2, PARK2, RNF187, CircXRN2 and various microRNAs [3,5,6,7,8].
How does negative regulation of hippo signaling affect cancer?
It can promote cancer by increasing YAP/TAZ nuclear activity, as seen in breast, bladder and esophageal cancers [3,5,6,7].
What diseases are linked to negative regulation of hippo signaling?
Breast cancer, bladder cancer, esophageal squamous carcinoma and musculoskeletal disorders have been linked to Hippo pathway dysregulation [1,3,5,6].
How can I study negative regulation of hippo signaling in the lab?
CRISPR knockout, point mutation, knock-in, overexpression, RNA-seq, proteomics and imaging are common approaches.
What is the role of YAP/TAZ in negative regulation of hippo signaling?
YAP/TAZ are downstream effectors; their nuclear localization indicates reduced Hippo signaling.
Can CRISPR be used to study negative regulation of hippo signaling?
Yes, CRISPR knockout, point mutation, knock-in and overexpression are widely used to test causal roles of genes in this process.
What is the relationship between microRNAs and negative regulation of hippo signaling?
MicroRNAs reciprocally regulate Hippo-YAP/TAZ signaling in breast cancer, influencing precision medicine approaches.
How does circXRN2 affect negative regulation of hippo signaling?
CircXRN2 suppresses tumor progression by activating the Hippo pathway in bladder cancer, counteracting negative regulation.
Conclusion
GO:0035331 negative regulation of hippo signaling is a critical biological process that controls YAP/TAZ activity and downstream growth programs. Its dysregulation is implicated in multiple cancers and musculoskeletal disorders, making it a rich area for mechanistic and translational research [1,3,5,6]. CRISPR-based models and functional screens are powerful tools to identify and validate negative regulators of Hippo signaling, and EDITGENE provides end-to-end support for such studies.
References
- 1. Han J et al.. 2024. Emerging role and function of Hippo-YAP/TAZ signaling pathway in musculoskeletal disorders.. Stem Cell Res Ther 15(1):386 PMID: 39468616
- 2. Tyrina E et al.. 2024. Hippo Signaling Pathway Involvement in Osteopotential Regulation of Murine Bone Marrow Cells Under Simulated Microgravity.. Cells 13(22) PMID: 39594669
- 3. Sadri F et al.. 2024. Hippo-YAP/TAZ signaling in breast cancer: Reciprocal regulation of microRNAs and implications in precision medicine.. Genes Dis 11(2):760-771 PMID: 37692482
- 4. Balavaishnavi B et al.. 2024. Regulation of hippo signaling mediated apoptosis by Rauvolfia tetraphylla in triple-negative breast cancer.. Med Oncol 41(5):103 PMID: 38553593
- 5. Xie B et al.. 2023. CircXRN2 suppresses tumor progression driven by histone lactylation through activating the Hippo pathway in human bladder cancer.. Mol Cancer 22(1):151 PMID: 37684641
- 6. Zhou X et al.. 2020. Regulation of Hippo/YAP signaling and Esophageal Squamous Carcinoma progression by an E3 ubiquitin ligase PARK2.. Theranostics 10(21):9443-9457 PMID: 32863938
- 7. Wang Z et al.. 2020. Regulation of Hippo signaling and triple negative breast cancer progression by an ubiquitin ligase RNF187.. Oncogenesis 9(3):36 PMID: 32198343
- 8. Wang L et al.. 2022. Multiphase coalescence mediates Hippo pathway activation.. Cell 185(23):4376-4393.e18 PMID: 36318920
