PAK4 methylation by the methyltransferase SETD6 attenuates cell adhesion.

PAK4 methylation by the methyltransferase SETD6 attenuates cell adhesion.

Scientific reports (2020-10-15)

Zlata Vershinin, Michal Feldman, Dan Levy

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.

MATERIALS

Product Number

Brand

Product Description

D5671

Sigma-Aldrich

Dulbecco′s Modified Eagle′s Medium - high glucose, With 4500 mg/L glucose and sodium bicarbonate, without L-glutamine and sodium pyruvate, liquid, sterile-filtered, suitable for cell culture, suitable for hybridoma

F1804

Sigma-Aldrich

Monoclonal ANTI-FLAG^® M2 antibody produced in mouse, 1 mg/mL, clone M2, affinity isolated antibody, buffered aqueous solution (50% glycerol, 10 mM sodium phosphate, and 150 mM NaCl, pH 7.4)

G7513

Sigma-Aldrich

L-Glutamine solution, 200 mM, solution, sterile-filtered, BioXtra, suitable for cell culture

P0781

Sigma-Aldrich

Penicillin-Streptomycin, with 10,000 units penicillin and 10 mg streptomycin per mL in 0.9% NaCl, 0.1 μm filtered, BioReagent, suitable for cell culture

05-904

Sigma-Aldrich

Anti-HA Tag Antibody, Upstate^®, from mouse

A2220

Millipore

ANTI-FLAG^® M2 Affinity Gel, purified immunoglobulin, buffered aqueous glycerol solution

M7145

Sigma-Aldrich

MEM Non-essential Amino Acid Solution (100×), without L-glutamine, liquid, sterile-filtered, BioReagent, suitable for cell culture