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- W2793409096 abstract "N-WASP is a potent activator of the Arp2/3 complex in vitro and harbours key regulatory functions for cellular actin assembly in response to signalling. To analyse N-WASP function, mice carrying a loxP flanked N-WASP allele were generated using gene targeting in embryonic stem cells and the Cre/loxP recombination system. N-WASPflox/flox mice allow the conditional inactivation of the N-WASP gene by Cre recombinase mediated deletion in a tissue specific and temporally controlled manner. The N-WASP gene was disrupted in the mouse germline, which led to embryonic lethality. To elucidate the role of N-WASP at the cellular level, embryonic fibroblasts from N-WASPflox/flox mice were immortalized and various N-WASP-defective cell lines were selected upon transient expression of Cre recombinase. These N-WASP-defective fibroblasts showed no apparent morphological alterations and were highly responsive to the induction of filopodia by microinjection of constitutively active Cdc42, but failed to support the intracellular motility of Shigella flexneri. Reconstitution experiments using GFP-tagged N-WASP and various GFP-N-WASP mutants revealed a recruitment mechanism independent of the CRIB domain and Cdc42. In addition, enteropathogenic and enterohemorrhagic Escherichia coli were incapable of inducing the formation of actin pedestals in N-WASP-defective cells. In reconstitution experiments, N-WASP recruitment and EPEC pedestal formation was shown to be independent of small Rho family GTPases and the CRIB domain, but to be mediated by a recruitment domain in the amino-terminal half of N-WASP. These results prove the essential role of N-WASP for actin cytoskeletal changes induced by these bacterial pathogens in vivo and in addition show for the first time that N-WASP is dispensable for filopodia formation." @default.
- W2793409096 created "2018-03-29" @default.
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- W2793409096 date "2002-04-30" @default.
- W2793409096 modified "2023-09-23" @default.
- W2793409096 title "Conditional Inactivation of N-WASP in the mouse: Analyses of N-WASP function" @default.
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- W2793409096 doi "https://doi.org/10.24355/dbbs.084-200511080100-650" @default.
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