Stoichiometry of Nck-dependent actin polymerization in living cells

Regulation of actin dynamics through the Nck/N-WASp (neural Wiskott-Aldrich syndrome protein)/Arp2/3 pathway is essential for organogenesis, cell invasiveness, and pathogen infection. Although many of the proteins involved in this pathway are known, the detailed mechanism by which it functions remai...

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Published in:The Journal of cell biology 2012-05, Vol.197 (5), p.643-658
Main Authors: Ditlev, Jonathon A, Michalski, Paul J, Huber, Greg, Rivera, Gonzalo M, Mohler, William A, Loew, Leslie M, Mayer, Bruce J
Format: Article
Language:English
Subjects:
Actins - chemistry
Actins - metabolism
Adaptor Proteins, Signal Transducing - chemistry
Adaptor Proteins, Signal Transducing - metabolism
Cell Survival
Cellular biology
Computer Simulation
HEK293 Cells
Humans
Oncogene Proteins - chemistry
Oncogene Proteins - metabolism
Pathogenesis
Polymerization
Proteins
Signal Transduction
Simulation
src Homology Domains
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cited_by cdi_FETCH-LOGICAL-c481t-9a4e3aeea504450f07842727d88381f33e479e742f988560e56e1463289fdf8c3
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container_title The Journal of cell biology
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creator Ditlev, Jonathon A
Michalski, Paul J
Huber, Greg
Rivera, Gonzalo M
Mohler, William A
Loew, Leslie M
Mayer, Bruce J
description Regulation of actin dynamics through the Nck/N-WASp (neural Wiskott-Aldrich syndrome protein)/Arp2/3 pathway is essential for organogenesis, cell invasiveness, and pathogen infection. Although many of the proteins involved in this pathway are known, the detailed mechanism by which it functions remains undetermined. To examine the signaling mechanism, we used a two-pronged strategy involving computational modeling and quantitative experimentation. We developed predictions for Nck-dependent actin polymerization using the Virtual Cell software system. In addition, we used antibody-induced aggregation of membrane-targeted Nck SH3 domains to test these predictions and to determine how the number of molecules in Nck aggregates and the density of aggregates affected localized actin polymerization in living cells. Our results indicate that the density of Nck molecules in aggregates is a critical determinant of actin polymerization. Furthermore, results from both computational simulations and experimentation support a model in which the Nck/N-WASp/Arp2/3 stoichiometry is 4:2:1. These results provide new insight into activities involving localized actin polymerization, including tumor cell invasion, microbial pathogenesis, and T cell activation.
doi_str_mv 10.1083/jcb.201111113
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subjects Actins - chemistry
Actins - metabolism
Adaptor Proteins, Signal Transducing - chemistry
Adaptor Proteins, Signal Transducing - metabolism
Cell Survival
Cellular biology
Computer Simulation
HEK293 Cells
Humans
Oncogene Proteins - chemistry
Oncogene Proteins - metabolism
Pathogenesis
Polymerization
Proteins
Signal Transduction
Simulation
src Homology Domains
title Stoichiometry of Nck-dependent actin polymerization in living cells
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