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A Yeast Genomic Screen to Identify Effectors of Kir2.1 Plasma Membrane Residence

Kolb, Alexander (2013) A Yeast Genomic Screen to Identify Effectors of Kir2.1 Plasma Membrane Residence. Doctoral Dissertation, University of Pittsburgh. (Unpublished)

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Protein quality control is important for the disposal of toxic misfolded proteins. Ion channels represent a class of proteins that are suited for the study of protein quality control within the secretory pathway. The inward rectifying potassium channel, Kir2.1, functions at the plasma membrane in several tissues to maintain membrane potential and osmotic homeostasis, and not surprisingly defects in Kir2.1 lead to a host of diseases. To identify Kir2.1 regulators, a yeast Kir2.1 expression system was established. Although a small population appeared to localize to the plasma membrane, Kir2.1 primarily localized to the ER and a significant fraction was targeted for ER associated-degradation (ERAD). To identify the spectrum of factors that maintain Kir2.1 plasma membrane residence, the expression plasmid was introduced into a potassium transporter deficient strain in which Kir2.1 restores growth on low potassium, and the strain was crossed to the deletion collection to obtain haploid progeny. Among the mutated genes that significantly increased cell growth on low potassium were those encoding members of the Endosomal Sorting Complex Required for Transport (ESCRT). ESCRT also regulated Kir2.1 stability in HeLa cells. These data indicate that Kir2.1 function is controlled by later steps in secretory pathway quality control, but not ERAD. The screen described represents a model that can be used for other ion channels.


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Item Type: University of Pittsburgh ETD
Status: Unpublished
CreatorsEmailPitt UsernameORCID
Kolb, Alexanderalk80@pitt.eduALK80
ETD Committee:
TitleMemberEmail AddressPitt UsernameORCID
Committee ChairBrodsky, Jeffrey Ljbrodsky@pitt.eduJBRODSKY
Committee MemberArndt, Karen Marndt@pitt.eduARNDT
Committee MemberTraub, Linton Mtraub@pitt.eduTRAUB
Committee MemberWeisz, Ora Aweisz@pitt.eduWEISZ
Committee MemberKleyman, Thomas Rkleyman@pitt.eduKLEYMAN
Date: 30 September 2013
Date Type: Publication
Defense Date: 15 April 2013
Approval Date: 30 September 2013
Submission Date: 17 June 2013
Access Restriction: 5 year -- Restrict access to University of Pittsburgh for a period of 5 years.
Number of Pages: 190
Institution: University of Pittsburgh
Schools and Programs: Dietrich School of Arts and Sciences > Biological Sciences
Degree: PhD - Doctor of Philosophy
Thesis Type: Doctoral Dissertation
Refereed: Yes
Uncontrolled Keywords: Protein Quality Control ESCRT Ion Channel Saccharomyces cerevisiae Protein Traffic TRK1
Date Deposited: 30 Sep 2013 11:53
Last Modified: 30 Sep 2018 05:15


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