Investigations Of The Functional Interface Between Env7 And Torc1 In Saccharomyces Cerevisiae
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Investigations Of The Functional Interface Between Env7 And Torc1 In Saccharomyces Cerevisiae
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Author : Jonathan Shaoulian
language : en
Publisher:
Release Date : 2016
Investigations Of The Functional Interface Between Env7 And Torc1 In Saccharomyces Cerevisiae written by Jonathan Shaoulian and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2016 with Autophagic vacuoles categories.
Abstract: In eukaryotic cells, deficiencies in proteins involved in lysosomal trafficking and function result in serious human disorders like Tay-Sach’s Disease or Alzheimer’s. The vacuole in Saccharomyces cerevisiae serves as a model for the mammalian lysosome. Our lab has identified ENV7, a novel gene involved in the late-endosome-to vacuole interface in Saccharomyces cerevisae. ENV7 (an ortholog of human STK16) encodes Env7, a vacuolar membrane kinase that negatively regulates vacuolar fusion. Tor1 is another conserved vacuolar membrane kinase, and has been proposed as a vacuolar fragmentation factor. Tor1, or its homolog Tor2, are the functional kinases of Tor Complex 1 (TORC1). Activated TORC1 stimulates cell growth and proliferation, as well as vacuolar fragmentation. In mammals, hyperactivity of TORC1 plays a major role in cancers and diabetes. Conversely, inhibition of TORC1 by starvation or treatment by the drug rapamycin, is associated with the initiation of autophagy, which itself involves a number of fusion events at the vacuole. Lastly, it has been shown that defects at various stages of vesicular trafficking play a role in disrupting Tor1 transport to the vacuole and perturbing the function of TORC1. Therefore, I hypothesized that there is a functional interface between Env7 and TORC1. Here I report that endogenously expressed Env7-GFP is localized to the vacuolar membrane, and upon inhibition of TORC1 by rapamycin, there is a significant mislocalization of Env7-GFP to diffuse cytoplasmic patterns. Overexpressed GFP-Env7 is also mislocalized when TORC1 activity is downregulated, but the mislocalization is to Golgi structures and to the plasma membrane. Additionally, while the absence of Tor1 is not sufficient to affect the localization of overexpressed GFP-Env7, when TORC1 is compromised by a deletion of TCO89 or by a more potent inhibition by rapamycin in both tor1[delta] and tco89[delta] cells, there is a significant mislocalization of GFP-Env7 to the plasma membrane. Taken together, my findings demonstrate that the mislocalization of Env7 is dependent on both the level of expression of Env7 and the state of TORC1 activity. Therefore, the results thus far support a possible functional interface between Env7 and TORC1. Furthermore, when TORC1 is inhibited by rapamycin treatment, unlike Env7-GFP, endogenously expressed Tor1-GFP remains on the vacuole, but forms numerous clusters at this location. In the course of testing my hypothesis, the effects of TORC1 inhibition on vacuolar morphology and onset of autophagy were also noted. I report that tor1[delta] cells, when treated by rapamycin, display a higher percentage of enlarged vacuole morphology relative to wt, and that this enlarged vacuole morphology seems to be associated with slightly increased levels of autophagy. This suggests that the absence of Tor1, and presumably its substitution by Tor2, compromises TORC1 and makes the cell more sensitive to rapamycin-induced autophagy. Additionally, I also report that absence of Env7 has no significant effect on vacuolar enlargement and the onset of autophagy in response to rapamycin treatment, and thus, does not affect the autophagy regulation function of TORC1. The results of this study suggest a functional interface between Env7 and TORC1. Since both are conserved in humans, such an interface may also be conserved in humans.
Psychosocial Framework Oxford Bibliographies Online Research Guide
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Author : Oxford University Press
language : en
Publisher: Oxford University Press, USA
Release Date : 2010-05-01
Psychosocial Framework Oxford Bibliographies Online Research Guide written by Oxford University Press and has been published by Oxford University Press, USA this book supported file pdf, txt, epub, kindle and other format this book has been release on 2010-05-01 with Social Science categories.
This ebook is a selective guide designed to help scholars and students of social work find reliable sources of information by directing them to the best available scholarly materials in whatever form or format they appear from books, chapters, and journal articles to online archives, electronic data sets, and blogs. Written by a leading international authority on the subject, the ebook provides bibliographic information supported by direct recommendations about which sources to consult and editorial commentary to make it clear how the cited sources are interrelated related. A reader will discover, for instance, the most reliable introductions and overviews to the topic, and the most important publications on various areas of scholarly interest within this topic. In social work, as in other disciplines, researchers at all levels are drowning in potentially useful scholarly information, and this guide has been created as a tool for cutting through that material to find the exact source you need. This ebook is a static version of an article from Oxford Bibliographies Online: Social Work, a dynamic, continuously updated, online resource designed to provide authoritative guidance through scholarship and other materials relevant to the study and practice of social work. Oxford Bibliographies Online covers most subject disciplines within the social science and humanities, for more information visit www.aboutobo.com.