The interaction between the adaptor protein APS and Enigma is involved in actin organisation

TY - JOUR

T1 - The interaction between the adaptor protein APS and Enigma is involved in actin organisation

AU - Barres, Romain

AU - Gonzalez, Teresa

AU - Le Marchand-Brustel, Yannick

AU - Tanti, Jean-François

PY - 2005/8/15

Y1 - 2005/8/15

N2 - APS (adaptor protein with PH and SH2 domains) is an adaptor protein phosphorylated by several tyrosine kinase receptors including the insulin receptor. To identify novel binding partners of APS, we performed yeast two-hybrid screening. We identified Enigma, a PDZ and LIM domain-containing protein that was previously shown to be associated with the actin cytoskeleton. In HEK 293 cells, Enigma interacted specifically with APS, but not with the APS-related protein SH2-B. This interaction required the NPTY motif of APS and the LIM domains of Enigma. In NIH-3T3 cells that express the insulin receptor, Enigma and APS were partially co-localised with F-actin in small ruffling structures. Insulin increased the complex formation between APS and Enigma and their co-localisation in large F-actin containing ruffles. While in NIH-3T3 and HeLa cells the co-expression of both Enigma and APS did not modify the actin cytoskeleton organisation, expression of Enigma alone led to the formation of F-actin clusters. Similar alteration in actin cytoskeleton organisation was observed in cells expressing both Enigma and APS with a mutation in the NPTY motif. These results identify Enigma as a novel APS-binding protein and suggest that the APS/Enigma complex plays a critical role in actin cytoskeleton organisation.

AB - APS (adaptor protein with PH and SH2 domains) is an adaptor protein phosphorylated by several tyrosine kinase receptors including the insulin receptor. To identify novel binding partners of APS, we performed yeast two-hybrid screening. We identified Enigma, a PDZ and LIM domain-containing protein that was previously shown to be associated with the actin cytoskeleton. In HEK 293 cells, Enigma interacted specifically with APS, but not with the APS-related protein SH2-B. This interaction required the NPTY motif of APS and the LIM domains of Enigma. In NIH-3T3 cells that express the insulin receptor, Enigma and APS were partially co-localised with F-actin in small ruffling structures. Insulin increased the complex formation between APS and Enigma and their co-localisation in large F-actin containing ruffles. While in NIH-3T3 and HeLa cells the co-expression of both Enigma and APS did not modify the actin cytoskeleton organisation, expression of Enigma alone led to the formation of F-actin clusters. Similar alteration in actin cytoskeleton organisation was observed in cells expressing both Enigma and APS with a mutation in the NPTY motif. These results identify Enigma as a novel APS-binding protein and suggest that the APS/Enigma complex plays a critical role in actin cytoskeleton organisation.

KW - Actin Cytoskeleton

KW - Actins

KW - Adaptor Proteins, Signal Transducing

KW - Amino Acid Motifs

KW - Animals

KW - Cell Differentiation

KW - Cell Surface Extensions

KW - Cytoskeletal Proteins

KW - Cytoskeleton

KW - HeLa Cells

KW - Humans

KW - Intracellular Signaling Peptides and Proteins

KW - LIM Domain Proteins

KW - Mice

KW - NIH 3T3 Cells

KW - Protein Binding

KW - Protein Structure, Tertiary

KW - Receptor, Insulin

U2 - 10.1016/j.yexcr.2005.05.008

DO - 10.1016/j.yexcr.2005.05.008

M3 - Journal article

C2 - 15946664

SN - 0014-4827

VL - 308

SP - 334

EP - 344

JO - Experimental Cell Research

JF - Experimental Cell Research

IS - 2

ER -