SURF1 | Leigh Syndrome

Keighley Reisenauer

[4]

What are mitochondrial disorder symptoms?

Mitochondrial disorders as windows into an ancient organelle, 2012: Nature

What is Leigh
Syndrome?
LS

Normal

Healthy, cute kid

http://www.amdf.org.au/dions-legacy/
Alaskan Husky Encephalopathy (AHE) Clinical Study, 2002: UC Davis
Immune-modulating drug unexpectedly benefits mice with fatal mitochondrial
defect. 2014, Univ. Washington

How are mitochondria affected in Leigh Syndrome?

[2]
http://www.ehealthzine.com/leigh-syndrome-causes.html

What gene is mutated in Leigh Syndrome?

SURF1

330

How to choose a model for neurodegeneration?

Citation

How well is SURF1 conserved between homologs?

114

330
283

67
68
77
79

293

337
289

74
89
78

284

311
353

What makes D. melanogaster a good model

organism?

Droso4Schools

What makes D. melanogaster a good model

organism?

Droso4Schools

What is the gap in knowledge?

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

Establish time of protein decay using SILAC

quantitative proteomics

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

How do ROS cause neuronal cell death?

http://sphweb.bumc.bu.edu/otl
t/MPHModules/PH/Aging/mobile_pa
ges/Aging3.html
www.sens.org

What is the ROS mechanism in

mitochondria?

www.wredenberglab.com

What is the ROS mechanism in

mitochondria?

e-

www.wredenberglab.com

What is the ROS mechanism in

mitochondria?

ROS

How can we measure ROS in mitochondria?

Esposti, Mauro Degli. Measuring mitochondrial reactive oxygen species. University of

Manchester, Science Direct Vol 46. 335-40

How can we measure ROS in mitochondria?

wellplate.com

How can we measure ROS in mitochondria?

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

SURF1 Relative Abundance

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/

SURF1 Relative Abundance

Final Conclusions

SURF1
Abundance
Overall

Time
SURF1 MT
Abundance

ROS
Abundance