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EXOCYST COMPLEX AND MEMBRANE TRAFFICKING IN POLARIZED EPITHELIAL CELLS

Oztan Matos, Asli (2008) EXOCYST COMPLEX AND MEMBRANE TRAFFICKING IN POLARIZED EPITHELIAL CELLS. Doctoral Dissertation, University of Pittsburgh. (Unpublished)

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Abstract

The octameric exocyst complex is associated with the junctional complex and recycling endosomes, and is proposed to selectively tether cargo vesicles directed toward the basolateral surface of polarized Madin-Darby canine kidney (MDCK) cells. I observed that the exocyst subunits Sec6, Sec8, and Exo70 were localized to early endosomes, transferrin-positive common recycling endosomes, and Rab11a-positive apical recycling endosomes of polarized MDCK cells. Consistent with its localization to multiple populations of endosomes, addition of function-blocking Sec8 antibodies to streptolysin-O permeabilized cells revealed exocyst requirements for several endocytic pathways including basolateral recycling, apical recycling, and basolateral-to-apical transcytosis. The latter was selectively dependent on interactions between the small GTPase Rab11a and Sec15A and was inhibited by the expression of the C-terminus of Sec15A or downregulation of Sec15A expression using shRNA. These results indicate that the exocyst complex may be a multi-purpose regulator of endocytic traffic directed toward both poles of polarized epithelial cells, and that transcytotic traffic is likely to require Rab11a-dependent recruitment and modulation of exocyst function, likely through interactions with Sec15A.


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Details

Item Type: University of Pittsburgh ETD
Status: Unpublished
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Oztan Matos, Asliasost2@pitt.eduASOST2
ETD Committee:
TitleMemberEmail AddressPitt UsernameORCID
Committee ChairTraub, Lintontraub@pitt.eduTRAUB
Committee MemberLinstedt, Adamlinstedt@andrew.cmu.edu
Committee MemberApodaca, Gerardgla6@pitt.eduGLA6
Committee MemberAridor, Meiraridor@pitt.eduARIDOR
Committee MemberWeisz, Oraweisz@msx.dept-med.pitt.eduWEISZ
Date: 14 February 2008
Date Type: Completion
Defense Date: 20 November 2007
Approval Date: 14 February 2008
Submission Date: 22 January 2008
Access Restriction: No restriction; Release the ETD for access worldwide immediately.
Institution: University of Pittsburgh
Schools and Programs: School of Medicine > Cell Biology and Molecular Physiology
Degree: PhD - Doctor of Philosophy
Thesis Type: Doctoral Dissertation
Refereed: Yes
Uncontrolled Keywords: apical; basolateral; exocyst; MDCK; membrane trafficking; polarized
Other ID: http://etd.library.pitt.edu/ETD/available/etd-01222008-164016/, etd-01222008-164016
Date Deposited: 10 Nov 2011 19:31
Last Modified: 15 Nov 2016 13:36
URI: http://d-scholarship.pitt.edu/id/eprint/6300

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