DTA

Digital Theses Archive

 

Tesi etd-12212021-135856

Type of thesis
Dottorato
Author
LAVILLA, MIKEL
URN
etd-12212021-135856
Title
A platform in yeast to understand oxygen sensing mechanisms in higher eukaryotes
Scientific disciplinary sector
BIO/04
Course
Istituto di Scienze della Vita - AGROBIOSCIENCES
Committee
relatore Prof. PERATA, PIERDOMENICO
Keywords
  • Oxygen
  • Plants
  • Hypoxia
  • Yeast
  • N-degron pathway
  • Synthetic Biology
  • Chemical Genetics
  • Transcriptional System
  • ERF-VII
Exam session start date
17/03/2022;
Availability
parziale
Abstract
Oxygen perception is essential to most terrestrial organisms. It allows them to switch from aerobic to anaerobic metabolism. Creating an orthogonal oxygen sensing device was achieved by transferring essential modules among species. In this project, we studied the expression of cysteine dioxygenases from plants (PLANT CYSTEINE OXIDASES, PCO) or animals (2-AMINOETHANETHIOL DIOXYGENASE, ADO) in the yeast Saccharomyces cerevisiae. These enzymes grant yeast the ability to confer an oxygen-conditional degradability to a bioluminescent protein module containing a Cys-exposing N-degron from RAP2.12, a well-characterized N-degron from plants involved in oxygen perception. Previously, we had reported the use of the Double Luciferase Oxygen Reporter (DLOR) for the quantitative analysis of oxygen dynamics in yeast. In this work, we present the use of this system to: (1) Characterize the effects of PCO point-mutations in its activity towards the degradation of RAP2.12, (2) Create a functional, high-throughput assay to define chemicals that may affect oxygen perception in plants and (3) Obtain a platform to test candidate proteins to be degraded through the ADO-mediated Cys/Arg N-degron pathway.
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