Clathrin-Mediated endocytosis, post-endocytic trafficking and their regulatory controls in plants (Record no. 374028)

000 -LEADER
fixed length control field 04369ntm a22003017a 4500
003 - CONTROL NUMBER IDENTIFIER
control field AT-ISTA
005 - DATE AND TIME OF LATEST TRANSACTION
control field 20190822113206.0
008 - FIXED-LENGTH DATA ELEMENTS--GENERAL INFORMATION
fixed length control field 190822s2019 au ||||| m||| 00| 0 eng d
040 ## - CATALOGING SOURCE
Transcribing agency IST
100 ## - MAIN ENTRY--PERSONAL NAME
Personal name Narasimhan, Madhumitha
9 (RLIN) 5912
245 ## - TITLE STATEMENT
Title Clathrin-Mediated endocytosis, post-endocytic trafficking and their regulatory controls in plants
260 ## - PUBLICATION, DISTRIBUTION, ETC. (IMPRINT)
Name of publisher, distributor, etc. IST Austria
Date of publication, distribution, etc. 2019
500 ## - GENERAL NOTE
General note Thesis
505 ## - FORMATTED CONTENTS NOTE
Formatted contents note Abstract
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Formatted contents note Acknowledgements
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Formatted contents note List of Figures
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Formatted contents note List of Tables
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Formatted contents note List of Symbols/Abbreviations
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Formatted contents note 1 Introduction
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Formatted contents note 2 A Functional Study of AUXILIN-LIKE1 and 2, Two Putative Clathrin Uncoating Factors in Arabidopsis
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Formatted contents note 3 Dissecting Clathrin-Coated Vesicle formation and trafficking in planta
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Formatted contents note 4 On deciphering the role of auxin on endocytotic regulation
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Formatted contents note 5 Conclusion and future prospects
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Formatted contents note 6 References
520 ## - SUMMARY, ETC.
Summary, etc. Clathrin-Mediated Endocytosis (CME) is an aspect of cellular trafficking that is constantly regulated for mediating developmental and physiological responses. The main aim of my thesis is to decipher the basic mechanisms of CME and post-endocytic trafficking in the whole multicellular organ systems of Arabidopsis. The first chapter of my thesis describes the search for new components involved in CME. Tandem affinity purification was conducted using CLC and its interacting partners were identified. Amongst the identified proteins were the Auxilin-likes1 and 2 (Axl1/2), putative uncoating factors, for which we made a full functional analysis. Over-expression of Axl1/2 causes extreme modifications in the dynamics of the machinery proteins and inhibition of endocytosis altogether. However the loss of function of the axl1/2 did not present any cellular or physiological phenotype, meaning Auxilin-likes do not form the major uncoating machinery. The second chapter of my thesis describes the establishment/utilisation of techniques to capture the dynamicity and the complexity of CME and post-endocytic trafficking. We have studied the development of endocytic pits at the PM – specifically, the mode of membrane remodeling during pit development and the role of actin in it, given plant cells possess high turgor pressure. Utilizing the improved z-resolution of TIRF and VAEM techniques, we captured the time-lapse of the endocytic events at the plasma membrane; and using particle detection software, we quantitatively analysed all the endocytic trajectories in an unbiased way to obtain the endocytic rate of the system. This together with the direct analysis of cargo internalisation from the PM provided an estimate on the endocytic potential of the cell. We also developed a methodology for ultrastructural analysis of different populations of Clathrin-Coated Structures (CCSs) in both PM and endomembranes in unroofed protoplasts. Structural analysis, together with the intensity profile of CCSs at the PM show that the mode of CCP development at the PM follows ‘Constant curvature model’; meaning that clathrin polymerisation energy is a major contributing factor of membrane remodeling. In addition, other analyses clearly show that actin is not required for membrane remodeling during invagination or any other step of CCP development, despite the prevalent high turgor pressure. However, actin is essential in orchestrating the post-endocytic trafficking of CCVs facilitating the EE formation. We also observed that the uncoating process post-endocytosis is not immediate; an alternative mechanism of uncoating – Sequential multi-step process – functions in the cell. Finally we also looked at one of the important physiological stimuli modulating the process – hormone, auxin. auxin has been known to influence CME before. We have made a detailed study on the concentration-time based effect of auxin on the machinery proteins, CCP development, and the specificity of cargoes endocytosed. To this end, we saw no general effect of auxin on CME at earlier time points. However, very low concentration of IAA, such as 50nM, accelerates endocytosis of specifically PIN2 through CME. Such a tight regulatory control with high specificity to PIN2 could be essential in modulating its polarity.
856 ## - ELECTRONIC LOCATION AND ACCESS
Uniform Resource Identifier <a href="https://doi.org/10.15479/at:ista:th1075">https://doi.org/10.15479/at:ista:th1075</a>
942 ## - ADDED ENTRY ELEMENTS (KOHA)
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Withdrawn status Lost status Source of classification or shelving scheme Damaged status Not for loan Permanent Location Current Location Date acquired Barcode Date last seen Price effective from Koha item type
  Not Lost       Library Library 2019-08-22 AT-ISTA#001884 2019-08-22 2019-08-22 Book

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