ARK5 AMP-activated protein kinase and glucose transport

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DESCRIPTION (provided by applicant): The immediate goal of this project is to train the candidate in molecular techniques (e.g. construction of vectors, transfection of mammalian cells, and protein transduction) to more closely examine the roles of individual proteins in the stimulation of glucose transport by insulin and other stimuli that the candidate studied during his postdoctoral training. The candidate's career aim is to apply diabetes research to a wide range of models, including cultured myotubes, isolated skeletal muscle, and muscle in vivo. The candidate's training at Saint Louis University will include mentored laboratory research, attendance of and participation in local research seminars and national conferences, and formal coursework in molecular biology and the responsible conduct of research. Diabetes researchers at a neighboring institution, Washington University, will provide additional technical training, career guidance, and research consultation. The environment offers numerous opportunities for the candidate to attend research seminars and interact with established researchers. The candidate has shown that stimulation of AMP-activated protein kinase (AMPK) activity is associated with increased insulin sensitivity in skeletal muscle, and he proposes to examine the role of ARK5, a newly-described Akt-activated AMPK family member, in the regulation of glucose transport. Since Akt is stimulated by muscle contractions (as recently reported in the literature and shown in preliminary data) and insulin, the hypothesis for this proposal is that ARK5 contributes to cross-talk between contraction- and insulin- signaling pathways in the stimulation of glucose transport. The candidate's preliminary data shows that skeletal muscle contains ARK5. The specific aims are 1) to determine whether ARK5 is activated by muscle contractions, insulin, and AMPK-activating stimuli, 2) to determine in myotubes (by means of overexpression of ARK5, expression of a constitutively active ARK5, and expression of ARK5 that cannot be regulated by Akt) the role of ARK5 in insulin-independent glucose transport and insulin sensitivity in the absence and presence of AMPK-activating stimuli, and 3) to determine, by transduction of ARK5 forms into myotubes, the acute role of ARK5 in the insulin sensitivity induced by AMPK-activating stimuli. The proposed training will develop the candidate in his transition to an independent researcher and is critical to the candidate's long-term ability to contribute to diabetes research