The insulin receptor substrate protein of 53 kDa (IRSp53) is critically involved in the formation of filopodia and neurites through mechanisms that have only in part been clarified. Here, we investigated the role of the small scaffold protein LIN7, an interactor of IRSp53. We found that formation of actin-filled protrusions in neuronal NSC34 cells and neurites in neuroblastoma N2A depends on motifs mediating the LIN7:IRSp53 association, as both the coexpression of LIN7 with IRSp53 or the expression of the L27-IRSp53 chimera (a fusion protein between IRSp53 and the LIN7L27 domain for plasma membrane protein complexes association) prevented actin-deficient protrusions induced by overexpressed IRSp53, and enhanced the formation of actin-filled protrusions. The regulatory role of LIN7 in IRSp53-mediated extension of filopodia was demonstrated by live-cell imaging experiments in neuronal N2A cells. Moreover, LIN7 silencing prevented the extension of filopodia and neurites, induced by ectopic expression of IRSp53 or serum starvation, respectively in undifferentiated and differentiated N2A cells. The expression of full length IRSp53 or the LIN7ΔPDZ mutant lacking the domain for association with IRSp53 was unable to restore neuritogenesis in LIN7 silenced cells. Conversely, defective neuritogenesis could be rescued by the expression of RNAi-resistant full length LIN7 or chimeric L27-IRSp53. Finally, LIN7 silencing prevented the recruitment of IRSp53 in Triton X-100 insoluble complexes, otherwise occurring in differentiated cells. Collectively these data indicate that LIN7 is a novel regulator of IRSp53, and that their association is required to promote the formation of actin-dependent filopodia and neurites.
LIN7 regulates the filopodia and neurite promoting activity of IRSp53 / A. Crespi, I. Ferrari, P. Lonati, A. Disanza, D. Fornasari, G. Scita, V. Padovano, G. Pietrini. - In: JOURNAL OF CELL SCIENCE. - ISSN 0021-9533. - 125:19(2012 Oct 01), pp. 4543-4554.
|Titolo:||LIN7 regulates the filopodia and neurite promoting activity of IRSp53|
CRESPI, ARIANNA (Primo)
FERRARI, ILARIA (Secondo)
PADOVANO, VALERIA (Penultimo)
PIETRINI, GRAZIA (Ultimo)
|Parole Chiave:||LIN7-IRSp53 association ; Actin-deficient filopodia ; Filopodium dynamics ; Motoneuronal-like NSC34 cells ; Neuritogenesis ; Neuroblastoma N2A cells|
|Settore Scientifico Disciplinare:||Settore MED/04 - Patologia Generale|
|Data di pubblicazione:||1-ott-2012|
|Digital Object Identifier (DOI):||http://dx.doi.org/10.1242/jcs.106484|
|Appare nelle tipologie:||01 - Articolo su periodico|