Genetic Pathways of the Hypoxic

Rescue of Frataxin Deficiency in Yeasts

Keong Mu Jason Lim, Jason Pitt, Alan Herr, Matt Kaeberlein
Undergraduate Research Symposium
May 21, 2021
University of Washington
Background & Significance

https://pubs.rsc.org/en/content/articlelan
ding/2014/CP/c3cp54055c#!divAbstract

https://sites.uab.edu/thenapieralalab/

Mootha et. al (2019), Hypoxia Rescues Frataxin Loss by Restoring Iron Sulfur Cluster Biogenesis, Cell
Methods
Find genes with similar oxygen Mat a yfh1∆
response as yfh1∆   Create BY4743 sporulate Mat α yfh1∆
+
Mat a YFH1
(target genes: sod1, sod2, rai1, nsr1, Mat α YFH1
pdc1, ctt1)
Hypoxia (0.5% O2)
 Create diploids
Mat a yfh1∆ x Mat α target genes sporulate & Normoxia (21% O2)
Ex) x
dissect
Hyperoxia (100% O2)

Screen for synthetic Replica plate Grow double mutants in

lethal/rescue obtain double mutants glucose vs glycerol environment
Mat a or α yfh1∆/sod1∆
Mat a or α yfh1∆/SOD1
Mat a or α YFH1/so1∆
Mat a or α YFH1/SOD1 (WT)
 Results
0.5% O2 21% O2 100% O2

YEPG

yfh1∆-1

YEPD

YEPG
• Able to reproduce Mootha et. al’s results
yfh1∆-2
• Only WT viable for normoxic and hyperoxic
conditions YEPD
• yfh1∆  viable with glucose but not with glycerol
Conclusion & Future Directions

• Conduct more experiments with double mutant strains

• Screen for rescue and inhibition mutation in double mutants

• Mimic hypoxic rescue with pharmacological agents?

Acknowledgments
I wish to thank Jason Pitt and Dr. Kaeberlein for mentoring and
providing me research opportunities for the past 4 years. I would also
like to thank Dr. Alan Herr and Andy Chia for helping me design and
carry out my experiments. I also wish to thank Anthony Grillo, Michael
Kiflezghi, and Michael Muir for the advice in experimental setups.