University of Illinois Urbana-Champaign

Optogenetic regulation of protein activity in live cell

Mondal, Payel

Content Files
MONDAL-DISSERTATION-2020.pdf

Permalink

https://hdl.handle.net/2142/108280

Description

Title
Optogenetic regulation of protein activity in live cell
Author(s)
Mondal, Payel
Issue Date
2020-05-07
Director of Research (if dissertation) or Advisor (if thesis)
Zhang, Kai
Doctoral Committee Chair(s)
Zhang, Kai
Committee Member(s)
  • Chen, Lin-Feng
  • Chen, Jie
  • Kalsotra, Auinash
Department of Study
Biochemistry
Discipline
Biochemistry
Degree Granting Institution
University of Illinois at Urbana-Champaign
Degree Name
Ph.D.
Degree Level
Dissertation
Date of Ingest
2020-08-27T00:50:04Z
Keyword(s)
  • optogenetics
  • GLIMPSe
  • degron
  • protein degradation
  • MKP3
  • CA MEK
  • BRD4
  • SPOP
  • PC12
  • Sonic Hedgehog
  • Sufu
  • Gli
  • Cargo Trafficking
Abstract
Signaling pathways extensively crosstalk among each other and result in different cellular phenotypes depending on the dynamic profile of protein activity. Conventional genetic and pharmacological approaches such as gene overexpression, use of growth factors or inhibitors have helped us delineate interaction maps of signaling components, however these techniques provide limited means to determine contribution of a target protein for specific cellular phenotype. Therefore, to find out the molecular mechanism for a cellular outcome, there is an urgent need for a tool that can specifically activate or inactivate a protein of interest and study it’s role towards a specific cell fate. Optogenetic techniques utilize light to control protein functions with high spatial and temporal resolution. Here, I First present a generalizable light modulated protein stabilization system (GLIMPSe) that enables target-independent optogenetic control of protein activities and minimizes the systematic variation embedded within different photoactivatable proteins. GLIMPSe was applied to control light-mediated post-translational stabilization of two distinct classes of proteins, phosphatase and kinase with rapid kinetics response. Second, I discuss role of speckle-type POZ protein (SPOP) in cell differentiation inhibition in PC12 and primary rat hippocampus neuron cell. Next, I combined the GLIMPSe system with the Sufu protein, a SPOP phenocopy and sonic hedgehog (Shh) signaling pathway inhibitor and generated the GLIMPSe-Sufu system for optogenetic inhibition of Shh signaling pathway. Finally, I review the molecular machinery of cargo trafficking with emphasis on new optogenetic and optochemical experimental strategies that enable direct modulation of cargo trafficking in live cells.
Graduation Semester
2020-05
Type of Resource
Thesis
Permalink
http://hdl.handle.net/2142/108280
Copyright and License Information
Copyright 2020 Payel Mondal

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Graduate Dissertations and Theses at Illinois PRIMARY
Graduate Theses and Dissertations at Illinois
Dissertations and Theses - Biochemistry