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(Unpublished)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Unpublished |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