Introduction
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Go Terms
ABCD1 is a protein found in the peroxisome. To be more precise, it is found in the peroxisomal membrane. It is a transporter which allows long chain fatty acids to pass through. This allows the peroxisome to break the fatty acids down via beta oxidation
Symptoms
What is a peroxisome?
Peroxisome are organelles involved in beta oxidation of fatty acids and transfer hydrogen. They contain the transporters on their membranes which allow for the fatty acids to come inside.
Conservation of ABCD1
ABCD1 is conserved very well between species. The figure indicates that most mammals have both domains convserved. As we look into different types of species, such as reptiles or amphibians, conservation begins to go down. Tiger shark for example contains only one of the domains and that domain itself is much smaller than what is seen in humans and other model organisms.
What is the gap?
Although ALD is an X-linked disease, it can be seen in females. The symptoms are much more severe in females than they are in males. What is causing the increased severity in males is unsure. This could be due to hormonal differences in between the sexes. The levels of protein associated with myelin and peroxisome maintenance must be much lower in males. This could cause the higher levels of fatty acids in males.
Zebrafish as Model Organism
Zebrafish was chosen as the model organism of choice because it is very easy to visualizes myelin which is what is being degraded in this disease. Fatty acids builds up in ALD and being able to see the buildup differences between males and females is important. Since there are a lot of pex genes, zebrafish allow for easy visualization of peroxisomes as well.
Aim 1
I wanted to determine conserved sequences in ABCD1 that are important for Myelin maintenance between male and females. I will do this by using ensemble to find the fasta sequences of each model organisms. Then I will align them on clustal omega. In conjuction to this, I found sites of phosphorylation that yielded high scores. I will look at these sites to see whether there could be anything related to the sex of the species. I found one spot at S269 that might be the case. Mutant fish will be created with crispr I hypothesieze that Amino acid S269A of ABCD1 may be important for sex-specific defects since the hermaphrodite was the only with an amino acid change.
Aim 2
The next part was to identify necessary genes necessary for myelin maintenance in males and females. The wild type and mutants will be run through RNA seq. Tissue samples from the CNS will be used. Sex defects in myelin as well as peroxisomes will be noted. I hypothesize genes involved in myelin maintenance and peroxisome function will be downregulated to a greater extent in males.
Aim 3
For my last aim, i will identify male-specific protein that mediate peroxisome function of ABCD1 that are important for myelin preservation. I will use iTRAQ and label both wildtype and mutant male and female with isotopes. These will be broken down and put in the same vile. Mass spec will be done to visualize the amount of protein in each group. I hypothesize that male-specific proteins important for myelin maintenance will be identified by comparing Wild type and ABCD1 mutant males and females.
Future Directions
I think one of the main things to look at is how this disease correlates to the other 2 forms of this disease. If they are similar, then treatment options for this disease will be viable for both.
How sex plays a role in this will also be interesting to see. Although they were not in the PPI, I wonder how much of an influence estrogen or testosterone play in the severity of this disease.
As mentioned in the chemical genomics, slide there is yet to find a drug to treat this disease. Finding something that can help treat this disease effectively should be found soon.
How sex plays a role in this will also be interesting to see. Although they were not in the PPI, I wonder how much of an influence estrogen or testosterone play in the severity of this disease.
As mentioned in the chemical genomics, slide there is yet to find a drug to treat this disease. Finding something that can help treat this disease effectively should be found soon.
References
•Baker, A., Carrier, D. J., Schaedler, T., Waterham, H. R., van Roermund, C. W., & Theodoulou, F. L. (2015, October). Peroxisomal ABC transporters: functions and mechanism. Retrieved from https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4652930/Yan, Fang & Wang, Wenbo & Ying, Hui & Li, Hongyu & Chen, Jing & Xu, Chao. (2017). S149R, a novel mutation in the ABCD1 gene Causing X-linked adrenoleukodystrophy. Oncotarget. 8. 10.18632/oncotarget.20974.
•seqlopedia. (n.d.). Retrieved from https://rnaseq.uoregon.edu/https://www.jbc.org/content/277/42/40142.long
•(n.d.). Retrieved from http://www.cbs.dtu.dk/services/NetPhos/
•RNA Sequencing. (n.d.). Retrieved from https://www.illumina.com/techniques/sequencing/rna-sequencing.html
Image References
•https://cdn.britannica.com/02/114902-050-0D7352BF/cells-organelles-mitochondria-chloroplasts-endoplasmic-reticulum-lysosomes.jpg
•https://www.creative-proteomics.com/images/iTRAQ-Workflow.jpg
•https://www.sisweb.com/mstools/spectrum.htm?id=507246695699227
•https://dmm.biologists.org/content/dmm/7/7/915/F1.large.jpg
•https://www.researchgate.net/profile/Olga_Krysko/publication/44681222/figure/fig1/AS:341280044404739@1458378911355/Peroxisomes-in-zebraWsh-embryos-Cross-sections-of-normal-96-hpf-zebraWsh-embryos-after_Q320.jpg
•https://www.researchgate.net/profile/Tim_Czopka/publication/242652467/figure/fig1/AS:339476648874001@1457948948929/Dynamic-Change-in-Global-Myelination-in-the-Larval-Zebrafish-Spinal-Cord-A-Lateral.png
•https://g.foolcdn.com/editorial/images/569850/pills-and-bottle.jpg
•seqlopedia. (n.d.). Retrieved from https://rnaseq.uoregon.edu/https://www.jbc.org/content/277/42/40142.long
•(n.d.). Retrieved from http://www.cbs.dtu.dk/services/NetPhos/
•RNA Sequencing. (n.d.). Retrieved from https://www.illumina.com/techniques/sequencing/rna-sequencing.html
Image References
•https://cdn.britannica.com/02/114902-050-0D7352BF/cells-organelles-mitochondria-chloroplasts-endoplasmic-reticulum-lysosomes.jpg
•https://www.creative-proteomics.com/images/iTRAQ-Workflow.jpg
•https://www.sisweb.com/mstools/spectrum.htm?id=507246695699227
•https://dmm.biologists.org/content/dmm/7/7/915/F1.large.jpg
•https://www.researchgate.net/profile/Olga_Krysko/publication/44681222/figure/fig1/AS:341280044404739@1458378911355/Peroxisomes-in-zebraWsh-embryos-Cross-sections-of-normal-96-hpf-zebraWsh-embryos-after_Q320.jpg
•https://www.researchgate.net/profile/Tim_Czopka/publication/242652467/figure/fig1/AS:339476648874001@1457948948929/Dynamic-Change-in-Global-Myelination-in-the-Larval-Zebrafish-Spinal-Cord-A-Lateral.png
•https://g.foolcdn.com/editorial/images/569850/pills-and-bottle.jpg
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