Targeted wild-type and jerker espins reveal a novel, WH2-domain-dependent way to make actin bundles in cells

Authors:
Loomis PA, Kelly AE, Zheng L, Changyaleket B, Sekerkova G, Mugnaini E, Ferreira A, Mullins RD, Bartles JR
In:
Source: J Cell Sci
Publication Date: (2006)
Issue: 119(Pt 8): 1655-65
Research Area:
Neurobiology
Cells used in publication:
Neuron, hippo/cortical, rat
Species: rat
Tissue Origin: brain
Neuron, mesencephalic, rat
Species: rat
Tissue Origin: brain
Neuron, hippocampal, rat
Species: rat
Tissue Origin: brain
Platform:
Nucleofector® I/II/2b
Abstract
The espin actin-bundling proteins, which are the target of deafness mutations, are present in the parallel actin bundles of stereocilia and microvilli and appear to increase their steady-state length. Here, we report a new activity of the espins, one that depends on their enigmatic WH2 domain: the ability to assemble a large actin bundle when targeted to a specific subcellular location. This activity was observed for wild-type espins targeted to the centrosome in transfected neuronal cells and for jerker espins targeted to the nucleolus in a wide variety of transfected cells as a result of the frameshifted peptide introduced into the espin C-terminus by the jerker deafness mutation. This activity, which appears specific to espins, requires two espin F-actin-binding sites and the actin-monomer-binding activity of the espin WH2 domain, but can be mimicked by adding a WH2 domain to an unrelated actin-bundling protein, villin. Espins do not activate the Arp2/3 complex in vitro, and bundle assembly is not indicative of in-vitro nucleation activity. Our results suggest a novel way to build actin bundles at specific sites in cells.