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Published 10 April 2006. doi:10.1083/jcb.200509076
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 173, Number 1, 121-132
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Article

Formation of a WIP-, WASp-, actin-, and myosin IIA–containing multiprotein complex in activated NK cells and its alteration by KIR inhibitory signaling



Konrad Krzewski1, Xi Chen1, Jordan S. Orange2, and Jack L. Strominger1

1 Department of Molecular and Cellular Biology, Harvard University, Cambridge, MA 02138
2 Division of Immunology, University of Pennsylvania School of Medicine, Children's Hospital of Philadelphia, Philadelphia, PA 19104

Correspondence to Jack L. Strominger: jlstrom{at}fas.harvard.edu

The tumor natural killer (NK) cell line YTS was used to examine the cytoskeletal rearrangements required for cytolysis. A multiprotein complex weighing ~1.3 mD and consisting of WASp-interacting protein (WIP), Wiskott-Aldrich syndrome protein (WASp), actin, and myosin IIA that formed during NK cell activation was identified. After induction of an inhibitory signal, the recruitment of actin and myosin IIA to a constitutive WIP–WASp complex was greatly decreased. Both actin and myosin IIA were recruited to WIP in the absence of WASp. This recruitment correlated with increased WIP phosphorylation, which was mediated by PKC{theta}. Furthermore, the disruption of WIP expression by WIP RNA interference prevented the formation of this protein complex and led to almost complete inhibition of cytotoxic activity. Thus, the multiprotein complex is important for NK cell function, killer cell immunoglobulin-like receptor inhibitory signaling affects proteins involved in cytoskeletal rearrangements, and WIP plays a central role in the formation of the complex and in the regulation of NK cell activity.

Abbreviations used in this paper: ITIM, immunoreceptor tyrosine-based inhibition motif; KIR, killer cell immunoglobulin-like receptor; MHC, major histocompatibility class; MS, mass spectrometry; NK, natural killer; WASp, Wiskott-Aldrich syndrome protein; WIP, WASp-interacting protein.


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