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Ctdnep1 and Eps8L2 regulate dorsal actin cables for nuclear positioning during cell migration.

Current biology : CB (2021-02-11)
Francisco J Calero-Cuenca, Daniel S Osorio, Sofia Carvalho-Marques, Sreerama Chaitanya Sridhara, Luis M Oliveira, Yue Jiao, Jheimmy Diaz, Cátia S Janota, Bruno Cadot, Edgar R Gomes
ZUSAMMENFASSUNG

Cells actively position their nuclei within the cytoplasm for multiple cellular and physiological functions.1-3 Consequently, nuclear mispositioning is usually associated with cell dysfunction and disease, from muscular disorders to cancer metastasis.4-7 Different cell types position their nuclei away from the leading edge during cell migration.8-11 In migrating fibroblasts, nuclear positioning is driven by an actin retrograde flow originated at the leading edge that drives dorsal actin cables away from the leading edge. The dorsal actin cables connect to the nuclear envelope by the linker of nucleoskeleton and cytoskeleton (LINC) complex on transmembrane actin-associated nuclear (TAN) lines.12-14 Dorsal actin cables are required for the formation of TAN lines. How dorsal actin cables are organized to promote TAN lines formation is unknown. Here, we report a role for Ctdnep1/Dullard, a nuclear envelope phosphatase,15-22 and the actin regulator Eps8L223-25 on nuclear positioning and cell migration. We demonstrate that Ctdnep1 and Eps8L2 directly interact, and this interaction is important for nuclear positioning and cell migration. We also show that Ctdnep1 and Eps8L2 are involved in the formation and thickness of dorsal actin cables required for TAN lines engagement during nuclear movement. We propose that Ctdnep1-Eps8L2 interaction regulates dorsal actin cables for nuclear movement during cell migration.

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Sigma-Aldrich
Monoklonaler ANTI-FLAG® M2-Antikörper in Maus hergestellte Antikörper, 1 mg/mL, clone M2, affinity isolated antibody, buffered aqueous solution (50% glycerol, 10 mM sodium phosphate, and 150 mM NaCl, pH 7.4)
Roche
DNase I, rekombinant, RNase-frei, from bovine pancreas, expressed in Pichia pastoris
Roche
Dispase® II (neutrale Protease, Klasse II), lyophilized, from bacterial, Roche, pkg of 5 × 1 g
Sigma-Aldrich
Monoklonaler Anti-Vinculin-Antikörper in Maus hergestellte Antikörper, clone hVIN-1, ascites fluid
Sigma-Aldrich
Triton X-100, laboratory grade
Sigma-Aldrich
Oleoyl-L-α-lysophosphatidische Säure Natriumsalz, ≥98%, solid
Sigma-Aldrich
Anti-EPS8L2 antibody produced in rabbit, Prestige Antibodies® Powered by Atlas Antibodies, affinity isolated antibody, buffered aqueous glycerol solution