Keighley Reisenauer
[4]
What is Leigh
Syndrome?
LS
Normal
[2]
http://www.ehealthzine.com/leigh-syndrome-causes.html
SURF1
330
Citation
114
330
283
67
68
77
79
293
337
289
74
89
78
284
311
353
Droso4Schools
Droso4Schools
Huge energy
demand
Must meet
huge energy
demand
http://en.hdbuzz.net/148
Edens, Brittany, Nimrod Miller, and Yong-Chao Ma. Impaired Autophagy and Defective Mitochondrial Function:
Converging Paths on the Road to Motor Neuron Degeneration. Frontiers of Cellular Neuroscience: March 2016.
What is my hypothesis?
My primary objective is to determine how loss of SURF1
in mitochondria impacts neuronal function.
Clker.com
Patil, Komal B. Electron Transport Chain Steps Explained with
Diagram. Buzzle: January 2015
What is my hypothesis?
My primary objective is to determine how loss of SURF1
in mitochondria impacts neuronal function.
MT SURF1
Clker.com
Patil, Komal B. Electron Transport Chain Steps Explained with
Diagram. Buzzle: January 2015
Aim 1
Determine dynamics of SURF1 in neural cells
Hypothesis: Mutated SURF1 degradation will occur shortly
after entering the cytosol and before reaching the
mitochondria.
Approach: CRISPR-Cas9 GFP tags on WT and MT SURF1
68
GFP
SURF
284
Aim 1
Determine dynamics of SURF1 in neural cells
Time
Relative # SURF
t=0
Lots
t=0.5
Less
t=1
Much less
Aim 2
Establish time of protein decay using SILAC
quantitative proteomics
Hypothesis: SURF1 will decrease in abundance with age in LS
patients and cells will show an increase of cellular death effects
over time.
Approach: SILAC labeling of WT and MT SURF1 D.
melanogaster neuron biopsies
https://www.thermofisher.com/us/en/home/life-science/protein-biology/protein-biology-learning-center/proteinbiology-resource-library/pierce-protein-methods/quantitativeproteomics/_jcr_content/MainParsys/image_f2ec.img.jpg/
Aim 2
Time
Relative Abundance
Aim 3
Understand how neurons die in mitochondrial
diseases, like LS
Hypothesis: SURF1 mutations result in COX IV defects causing
an imbalance of the proton gradient, thus an increase of lethal
ROS, leading to neuronal cell death.
Approach: Measure ROS markers from Aim 2 biopsies
http://hmg.oxfordjournals.org/c
ontent/10/20/2277.full.pdf
http://sphweb.bumc.bu.edu/otl
t/MPHModules/PH/Aging/mobile_pa
ges/Aging3.html
www.sens.org
www.wredenberglab.com
e-
www.wredenberglab.com
ROS
wellplate.com
wellplate.com
Time
SURF1 Relative
Abundance
SURF1 Relative
Abundance
Conclusions
Time
SURF1 Relative
Abundance
Conclusions
Time
SURF1
Abundance
Overall
SURF1 MT
Abundance
Time
ROS Relative
Abundance
SURF1 Relative
Abundance
Conclusions
Time
Final Conclusions
SURF1
Abundance
Overall
Time
SURF1 MT
Abundance
ROS
Abundance
Future Directions
- Drug testing
- alpha-trocotrienol quinone (EPI-743)
Martinelli, Diego, et. al. "EPI-743 reverses the progression of the pediatric mitochondrial diseaseGenetically
defined Leigh Syndrome." Molecular Genetics and Metabolism: Volume 107, Issue 3. Science Direct. November
2012. 383-388.
References
1.
http://www.thelancet.com/journals/laneur/article/PIIS1474-4422(15)00148-9/fulltext?rss=yes
2.
http://journal.frontiersin.org/article/10.3389/fgene.2012.00244/full
3.
http://ghr.nlm.nih.gov/gene/SURF1
4.
https://www.youtube.com/watch?v=0-EB2kbLhAs
5.
http://www.nature.com/nrm/journal/v9/n7/box/nrm2434_BX1.html
6.
http://theawkwardyeti.com/tag/mitochondria/
7.
http://www.beautifulyou.com.pk/article/bust-the-myth/
Final Conclusions
SURF1
Abundance
Overall
Time
SURF1 MT
Abundance
ROS
Abundance