Ravenall, Samantha Jane; (2002) Cytoskeletal regulation by a novel peripheral nervous system-specific actin binding protein. Doctoral thesis (Ph.D), UCL (University College London).

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Abstract

The cytoskeleton plays an essential role in cell development, morphology and plasticity. In neurons, the cytoskeleton has been implicated in many functions including signal transduction, neurite outgrowth and transport of organelles. This thesis focuses on a putative cytoskeletal regulatory protein found to be expressed selectively in peripheral sensory and sympathetic neurons. The transcript encoding the villin-like protein was identified in a difference-cloning exercise to find sensory neuron-specific genes. Pervin, the predicted 829 amino acid protein, has 60% identity with villin and is the rat homologue of advillin. Immunoprecipitation studies show that pervin and actin interact together in vivo. Transfection of COS-7 fibroblast cell lines demonstrates co-localisation of epitope-tagged pervin with GFP-actin and results in an increase in process formation. This effect is abolished when the putative actin-bundling headpiece of pervin is deleted. Biolistic transfection of primary cultures of rat dorsal root ganglion sensory neurons results in increased neurite outgrowth with FLAG-tagged pervin. Deletion of the actin bundling headpiece inhibits normal neurite growth. These data suggest that pervin may play a significant role in regulating process outgrowth in peripheral neurons through a mechanism that involves the activity of an actin-bundling domain.

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