Identification of PASK substrates and/or interacting proteins

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Publication Type honors thesis
School or College College of Science
Department Biology
Faculty Mentor Jared Rutter
Creator Johnson, KariAnne Rencher
Title Identification of PASK substrates and/or interacting proteins
Year graduated 2012
Date 2012-05
Description PAS kinase (PASK) is an evolutionary conserved serine/threonine kinase that appears to have a role in the regulation of cellular energy metabolism. It has previously been shown that mice lacking PASK (PASK"/_ mice) are resistant to the development of hepatic steatosis (lipid accumulation in the liver), glucose intolerance, insulin resistance and obesity when placed on a high fat diet. These results suggest that PASK could be a potential target for the treatment of type II diabetes and obesity, However, the function of PASK at the cellular level is unknown. The aim of this project is to identify substrates and/or interacting partners for PASK in order to better understand its biological significance and connection to metabolic diseases. In order to achieve this objective, FLAG and HA dually-tagged constructs expressing wild type (WT) or kinase inactive (KI) PASK were generated in a viral expression vector. HepG2 cells were then infected with viral particles encoding Nterminally tagged WT or N-terminally tagged KI PASK. Tandem affinity purification (TAP) was optimized for this cell line. After purification, PASK-associated proteins were identified by mass spectrometry. Thyroid hormone receptor associated protein 3 (THRAP3), a protein involved in mRNA splicing, was identified as a potential interacting protein. However, subsequent experiments failed to reproduce this interaction with PASK in co-immunoprecipitation experiments.
Type Text
Publisher University of Utah
Subject Biology
Language eng
Rights Management (c) KariAnne Rencher Johnson
Format Medium application/pdf
Format Extent 732,456 bytes
Permissions Reference URL
ARK ark:/87278/s63r434k
Setname ir_htoa
Date Created 2016-10-31
Date Modified 2019-07-10
ID 205857
Reference URL
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