the adaptor molecule nck localizes the wave complex to promote actin polymerization during ceacam3-mediated phagocytosis of bacteria适配器分子nck所在波复杂促进肌动蛋白聚合在ceacam3-mediated吞噬的细菌.pdfVIP

The Adaptor Molecule Nck Localizes the WAVE Complex to Promote Actin Polymerization during CEACAM3- Mediated Phagocytosis of Bacteria 1. 1. 2.¤ ´ 1 1 Stefan Pils , Kathrin Kopp , Lisa Peterson , Julia Delgado Tascon , Naja J. Nyffenegger-Jann , Christof R. Hauck1,3* ¨ ¨ ¨ ¨ ¨ 1 Lehrstuhl Zellbiologie, Universitat Konstanz, Konstanz, Germany, 2 Zentrum fur Infektionsforschung, Universitat Wurzburg, Wurzburg, Germany, 3 Konstanz Research ¨ School Chemical Biology, Universitat Konstanz, Konstanz, Germany Abstract Background: CEACAM3 is a granulocyte receptor mediating the opsonin-independent recognition and phagocytosis of human-restricted CEACAM-binding bacteria. CEACAM3 function depends on an intracellular immunoreceptor tyrosine- based activation motif (ITAM)-like sequence that is tyrosine phosphorylated by Src family kinases upon receptor engagement. The phosphorylated ITAM-like sequence triggers GTP-loading of Rac by directly associating with the guanine nucleotide exchange factor (GEF) Vav. Rac stimulation in turn is critical for actin cytoskeleton rearrangements that generate lamellipodial protrusions and lead to bacterial uptake. Principal Findings: In our present study we provide biochemical and microscopic evidence that the adaptor proteins Nck1 and Nck2, but not CrkL, Grb2 or SLP-76, bind to tyrosine phosphorylated CEACAM3. The association is phosphorylation- dependent and requires the Nck SH2 domain. Overexpression of the isolated Nck1 SH2 domain, RNAi-mediated knock-down of Nck