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  • ZNF185 prevents stress fiber formation through the inhibition of RhoA in endothelial cells.

ZNF185 prevents stress fiber formation through the inhibition of RhoA in endothelial cells.

Communications biology (2023-01-12)
Soichiro Suzuki, Fumiaki Ando, Sae Kitagawa, Yu Hara, Tamami Fujiki, Shintaro Mandai, Koichiro Susa, Takayasu Mori, Eisei Sohara, Tatemitsu Rai, Shinichi Uchida
ABSTRACT

Signaling through cAMP/protein kinase A (PKA) promotes endothelial barrier function to prevent plasma leakage induced by inflammatory mediators. The discovery of PKA substrates in endothelial cells increases our understanding of the molecular mechanisms involved in vessel maturation. In this study, we evaluate a cAMP inducer, forskolin, and a phospho-PKA substrate antibody to identify ZNF185 as a PKA substrate. ZNF185 interacts with PKA and colocalizes with F-actin in endothelial cells. Both ZNF185 and F-actin accumulate in the plasma membrane region in response to forskolin to stabilize the cortical actin structure. By contrast, ZNF185 knockdown disrupts actin filaments and promotes stress fiber formation without inflammatory mediators. Constitutive activation of RhoA is induced by ZNF185 knockdown, which results in forskolin-resistant endothelial barrier dysfunction. Knockout of mouse Zfp185 which is an orthologous gene of human ZNF185 increases vascular leakage in response to inflammatory stimuli in vivo. Thrombin protease is used as a positive control to assemble stress fibers via RhoA activation. Unexpectedly, ZNF185 is cleaved by thrombin, resulting in an N-terminal actin-targeting domain and a C-terminal PKA-interacting domain. Irreversible dysfunction of ZNF185 protein potentially causes RhoA-dependent stress fiber formation by thrombin.

MATERIALS
Product Number
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Product Description

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