Understanding the Regulation of the Human Amino Acid Transporter ASCT2

Simple item page
dc.contributor.advisorDonini, Andrew
dc.contributor.advisorCoe, Imogen
dc.creatorPlastina, Maria Cristina
dc.date.accessioned2018-05-28T12:53:52Z
dc.date.available2018-05-28T12:53:52Z
dc.date.copyright2017-12-04
dc.date.issued2018-05-28
dc.date.updated2018-05-28T12:53:52Z
dc.degree.disciplineBiology
dc.degree.levelDoctoral
dc.degree.namePhD - Doctor of Philosophy
dc.description.abstractGlutamine is a very versatile source of energy and a precursor for many molecules which are essential to the growth and survival of actively dividing cells. Numerous amino acid transporters are involved in the transport and homeostasis of glutamine and ASCT2 is one of the most important transporters expressed in mammals, due to its moderate affinity for glutamine (Km = 70 - 90 M), its ubiquitous organ expression, and its correlation with aggressive forms of cancer. Yet, the regulation of ASCT2 in a non-pathological setting is poorly understood. In this study, I developed a new cell model for the study of ASCT2 by examining the molecular and functional characteristics of glutamine uptake in RWPE-1, an immortalized prostate epithelial human cell line. The expression of the canonical isoform of ASCT2 (SLC1A5) mRNA in RWPE-1 cells was confirmed by quantitative PCR and ASCT2 protein by western blotting. Immunofluorescence microscopy confirmed ASCT2 to be abundant and primarily localized to the plasma membrane. Radio-labelled glutamine uptake in RWPE-1 cells determined that ASCT2 is the major contributor (45 %) of glutamine transport in this cell line. Additionally, I have confirmed that EGF regulates the glutamine uptake via ASCT2, by regulating, through an unknown mechanism, the trafficking of the transporter to the plasma membrane. Thus, I conclude that the RWPE-1 cell-line is the first model to date used to study the detailed assessment of amino acid transporters role in the physiology of the prostate; moreover, RWPE-1 is a physiologically relevant model for the study of ASCT2 regulation. Moreover, I performed a largescale MYTH screening to target ASCT2 and discover potential interactors of the transporter which may help my future studies to further investigate and understand the regulation of the transporter.
dc.identifier.urihttp://hdl.handle.net/10315/34540
dc.language.isoen
dc.rightsAuthor owns copyright, except where explicitly noted. Please contact the author directly with licensing requests.
dc.subjectMolecular biology
dc.subject.keywordsASCT2
dc.subject.keywordsSLC1A5
dc.subject.keywordsRWPE-1
dc.subject.keywordsRegulation
dc.subject.keywordsEGF
dc.subject.keywordsMYTH
dc.subject.keywordsInteractome
dc.titleUnderstanding the Regulation of the Human Amino Acid Transporter ASCT2
dc.typeElectronic Thesis or Dissertation

Files

Original bundle
Now showing 1 - 1 of 1
Thumbnail Image
Name:
Plastina_MariaCristina_2017_PhD.pdf
Size:
3.8 MB
Format:
Adobe Portable Document Format
Download
License bundle
Now showing 1 - 2 of 2
No Thumbnail Available
Name:
license.txt
Size:
1.83 KB
Format:
Plain Text
Description:
Download
No Thumbnail Available
Name:
YorkU_ETDlicense.txt
Size:
3.38 KB
Format:
Plain Text
Description:
Download

Collections

Biology

All items in the YorkSpace institutional repository are protected by copyright, with all rights reserved except where explicitly noted.