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Title: Functional analysis of NET2A at the actin-membrane interface of plants
Author: Duckney, Patrick James
ISNI:       0000 0004 6350 4900
Awarding Body: Durham University
Current Institution: Durham University
Date of Award: 2017
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Recently, NET2A was characterised as a novel, plant-specific actin-binding protein that binds actin at the plasma membrane of growing pollen tubes (Deeks et al. 2012; Dixon. 2013). However, a function for NET2A has not yet been identified. Throughout the course of this investigation, several strategies were employed to elucidate the roles of NET2A in Arabidopsis, including reverse-genetic analysis, in situ localisation studies of NET2A in developing pollen grains and growing pollen tubes, as well as several protein-protein interaction screens. During this study, multiple NET2 subfamily members were shown to interact with plasma membrane integral proteins, demonstrating novel mechanisms by which the actin cytoskeleton is linked to the plasma membrane in plants. Through these interactions, NET2 proteins are implicated in the regulation of pollen tube growth in vivo during fertilisation, and regulation of the cytoskeleton in response to extracellular signals. A role for NET2A in pollen grain development was also studied during this investigation, in which NET2A was observed to associate with the actin cytoskeleton of developing pollen grains, and undergo dynamic reorganisations in subcellular localisation timed to specific developmental events in male gametogenesis. Reverse-genetic analysis of individual NET2 subfamily members has identified no phenotypic growth defects in pollen grain development or fertilisation. Several lines of evidence suggest functional redundancy and cooperation between individual NET2 proteins, including their involvement with common interacting partners at the plasma membrane, and interactions with one another in NET2 hetero-oligomers. Additionally, this project describes the discovery of a novel microtubule-associated protein in Arabidopsis. This protein, named herein as MAP7A, was demonstrated to associate with microtubules directly, and localise to the pollen tube plasma membrane and generative cell nucleus. Described in this thesis are the potential roles for NET2A and MAP7A in the regulation of the plant cytoskeleton during anisotropic cell growth.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available