The ultimate goal of my research is to understand molecular mechanisms underlying the subcellular localization of proteins in neurons. A more immediate goal is to use the recent discovery of a dendritic targeting motif in the C-terminus of the potassium channel Kv4.2 to understand specific molecular mechanisms involved in dendritic targeting of that channel. The research career development plan proposed in this grant involves five major components: Training in new techniques including patch clamping of transfected cells, patch clamping and intracellular recording of neurons in slices, and techniques of electron microscopy; Running a journal club; Attending conferences; Organizing a departmental retreat; Teaching in an ethics of research course. Studies conducted under this grant are aimed at understanding, in molecular terms, the mechanisms by which voltage-gated ion channels are targeted to specific subcellular domains of neurons. The proposed experiments are based on the hypothesis that subcellular targeting of neuronal transmembrane proteins is mediated by short amino acid sequences that interact with adaptor protein complexes which, in turn, bind directly or indirectly to kinesins. This molecular machinery is responsible for sorting proteins into vesicles and transporting the vesicles to specific subcellular locations. In previous work we identified a targeting signal encoded by a stretch of 16 amino acids within the C-terminus of Kv4.2 and showed that it is necessary and sufficient for dendritic localization. In this study we plan to extend these findings by determining the point in the targeting process at which the motif acts and which adaptor protein complex and kinesin isoform act downstream of the motif. These studies will enable us to understand how the presence of a short peptide signal on a protein can lead to the targeting of that protein to a specific subcellular region in neurons.
| Lewis Jr, Tommy L; Mao, Tianyi; Arnold, Don B (2011) A role for myosin VI in the localization of axonal proteins. PLoS Biol 9:e1001021 |
| Lewis Jr, Tommy L; Mao, Tianyi; Svoboda, Karel et al. (2009) Myosin-dependent targeting of transmembrane proteins to neuronal dendrites. Nat Neurosci 12:568-76 |
| Arnold, Don B (2009) Actin and microtubule-based cytoskeletal cues direct polarized targeting of proteins in neurons. Sci Signal 2:pe49 |
| Arnold, Don B (2007) Polarized targeting of ion channels in neurons. Pflugers Arch 453:763-9 |
| Rivera, Jacqueline; Chu, Po-Ju; Lewis Jr, Tommy L et al. (2007) The role of Kif5B in axonal localization of Kv1 K(+) channels. Eur J Neurosci 25:136-46 |
