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Updated: Dec 6, 2025
An Alternative Culture Method to Maintain Genomic Hypomethylation of Mouse Embryonic Stem Cells Using MEK Inhibitor PD0325901 and Vitamin C
Published on: June 1, 2018
PAK4 methylation by the methyltransferase SETD6 attenuates cell adhesion
Zlata Vershinin1,2, Michal Feldman1,2, Dan Levy3,4
1The Shraga Segal Department of Microbiology, Immunology and Genetics, Ben-Gurion University of the Negev, P.O.B. 653, 84105, Be'er-Sheva, Israel.
Abstract:
P21-activated kinase 4 (PAK4), a member of serine/threonine kinases family is over-expressed in numerous cancer tumors and is associated with oncogenic cell proliferation, migration and invasion. Our recent work demonstrated that the SET-domain containing protein 6 (SETD6) interacts with and methylates PAK4 at chromatin in mammalian cells, leading to activation of the Wnt/β-catenin signaling pathway. In our current work, we identified lysine 473 (K473) on PAK4 as the primary methylation site by SETD6. Methylation of PAK4 at K473 activates β-catenin transcriptional activity and inhibits cell adhesion. Specific methylation of PAK4 at K473 also attenuates paxillin localization to focal adhesions leading to overall reduction in adhesion-related features, such as filopodia and actin structures. The altered adhesion of the PAK4 wild-type cells is accompanied with a decrease in the migrative and invasive characteristics of the cells. Taken together, our results suggest that methylation of PAK4 at K473 plays a vital role in the regulation of cell adhesion and migration.
Insights
Methylation of P21-activated kinase 4 (PAK4) at lysine 473 by SETD6 activates Wnt/β-catenin signaling, inhibiting cell adhesion and migration. This finding reveals a key regulatory mechanism in cancer progression.
Area of Science:
- Molecular Biology
- Cell Biology
- Biochemistry
Background:
- P21-activated kinase 4 (PAK4) is a serine/threonine kinase implicated in cancer, promoting proliferation, migration, and invasion.
- SET-domain containing protein 6 (SETD6) has been shown to interact with and methylate PAK4, activating the Wnt/β-catenin signaling pathway.
Purpose of the Study:
- To identify the specific methylation site on PAK4 targeted by SETD6.
- To elucidate the functional consequences of PAK4 methylation at this site on cellular processes.
Main Methods:
- Site-directed mutagenesis to identify the primary methylation site (K473).
- Analysis of β-catenin transcriptional activity.
- Assessment of cell adhesion, paxillin localization, and actin structures.
- Evaluation of cell migration and invasion assays.
Main Results:
- Lysine 473 (K473) was identified as the primary site of PAK4 methylation by SETD6.
- Methylation of PAK4 at K473 enhances β-catenin transcriptional activity and reduces cell adhesion.
- PAK4 methylation at K473 disrupts paxillin localization to focal adhesions, affecting filopodia and actin organization.
- Reduced cell adhesion correlates with decreased cell migration and invasion.
Conclusions:
- Methylation of PAK4 at K473 is a critical regulatory event.
- This post-translational modification plays a significant role in modulating cell adhesion, migration, and invasion.
- The SETD6-mediated methylation of PAK4 offers a potential therapeutic target for cancers.
