Myo5a, Melanosome Transport and Lavender Foal Syndrome

Myo5a, Melanosome Transport and Lavender Foal Syndrome

Myo5a, Melanosome Transport and Lavender Foal Syndrome Rebecca Blackburn h=p://www.cellmigraDon.org/resource/discovery/vale What is Lavender Foal Syndrome? h=p://www.equinenow.com/farm/cressant_hill_arabians.htm h=p://arabiansltd.com/recent-­‐arabian-­‐sales What are the symptoms? h=p://delightarabhorsestud.homestead.com/Foals_.html h=p://animalgeneDcs.eu/Equine/equine-­‐geneDc-­‐disease/lfs.html Normal LFS h=p://livingmydreamkn.blogspot.com/2011/04 Divers, TJ. (2006) Structural and FuncDonal Diseases of the Equine Brain. Clinical Techniques in Equine prac1ce. 5(1): 67-­‐73 Normal LFS Myo5a mutaDons cause abnormal pigmentaDon Normal h=p://www.more4kids.info/699/childrens-­‐intuiDon/ Normal h=p://jaxmice.jax.org/jaxnotes/512/512l.html Myo5a-­‐ h=p://www.fcarreras.org/en/mohammed-­‐griscelli-­‐syndrome-­‐former-­‐paDent_42620 Myo5a-­‐ h=ps://database.riken.jp/sw/en/B10.D1-­‐Myo5a_d__Hir How does Myo5a funcDon in the cell? Kneussel, M. Wagner, W. (2013) Myosin motors and neuronal synapses: drivers of membrane transport and acDn dynamics. Nature Reviews: Neuroscience. 14: 233-­‐48. Myo5a is mutated in Lavender Foal Syndrome Myosin Head IQ Coiled Coil Low Complexity DIL Base DeleDon Kneussel, M. Wagner, W. (2013) Myosin motors and neuronal synapses: drivers of membrane transport and acDn dynamics. Nature Reviews: Neuroscience. 14: 233-­‐48. PhylogeneDc Analysis Myo5a is highly conserved Myosin Head IQ Coiled Coil Low Complexity DIL % Similarity 99% 97% 96% 98% 94% Myo5a is highly conserved Myosin Head IQ Coiled Coil Low Complexity DIL % Similarity 86% 58% 60% 55% 57% 56% Hypothesis: Low complexity region is important for dimerizaDon Myosin Head IQ Coiled Coil Low Complexity DIL Is the low complexity region conserved? Mutagenesis of Low Complexity Region Myosin Head IQ Coiled Coil Low Complexity DIL Mutagenesis = no dimerizaDon Myosin Head IQ Coiled Coil Low Complexity DIL Myosin Head IQ Coiled Coil Low Complexity DIL Assay for dimerizaDon – TAP tag Huber, L. A. (2003) Is proteomics heading in the wrong direcDon? Nature Reviews Molecular Cell Biology, 4, 74-­‐80 Myo5a mutaDons cause abnormal pigmentaDon Normal h=p://www.more4kids.info/699/childrens-­‐intuiDon/ Normal h=p://jaxmice.jax.org/jaxnotes/512/512l.html Myo5a-­‐ h=p://www.fcarreras.org/en/mohammed-­‐griscelli-­‐syndrome-­‐former-­‐paDent_42620 Myo5a-­‐ h=ps://database.riken.jp/sw/en/B10.D1-­‐Myo5a_d__Hir Myo5a transports melanosomes h=p://kaDya-­‐junkyard.blogspot.com/2013/03/physiology-­‐of-­‐color-­‐and-­‐pa=ern-­‐in.html Kuroda, T.S. et al. (2003). The acDn binding domain of slac-­‐2a/melanophilin is required for melanosome distribuDon in melanocytes. Mol Cel Biol. 23(15): 5245-­‐55. Hypothesis: There are unidenDfied proteins important for pigmentaDon that bind to Myo5a h=p://kaDya-­‐junkyard.blogspot.com/2013/03/physiology-­‐of-­‐color-­‐and-­‐pa=ern-­‐in.html Kuroda, T.S. et al. (2003). The acDn binding domain of slac-­‐2a/melanophilin is required for melanosome distribuDon in melanocytes. Mol Cel Biol. 23(15): 5245-­‐55. Why use zebra fish to study Melanosomes? h=p://post.jagran.com/sakhi-­‐Zebrafish-­‐to-­‐mend-­‐a-­‐broken-­‐heart-­‐1321348010 h=p://www.erasmusmc.nl/klinische_geneDca/research/intro/willemsen/
zebrafish/?lang=en Genome wide screen to idenDfy proteins involved in pigmentaDon h=p://kaDya-­‐junkyard.blogspot.com/2013/03/physiology-­‐of-­‐color-­‐and-­‐pa=ern-­‐in.html Luo, N. et al. (2012) Evidence for a role of inositol polyphosphate 5-­‐
phosphatase INPP5E in cilia formaDon in zebrafish. Vision Research. 75:98-­‐107. Sort proteins using gene ontology Vesicle Transport Zebra fish Protein Web Apoptosis Organelle Biogenesis DNA Repair h=p://string-­‐db.org/ Assay for Myo5a interacDon – TAP tag Huber, L. A. (2003) Is proteomics heading in the wrong direcDon? Nature Reviews Molecular Cell Biology, 4, 74-­‐80 Screen to look for novel transport proteins Control Luo, N. et al. (2012) Evidence for a role of inositol polyphosphate 5-­‐
phosphatase INPP5E in cilia formaDon in zebrafish. Vision Research. 75:98-­‐107. Expected Why study melanosome transport? Melanocyte Marks, MS. Seabra, MC. (2001) The Melanosome: Membrane Dynamics in Black and White. Nature Reviews: Molecular Cell Biology. 2:1-­‐11. Neuron Cai, Q. Davis, ML. Sheng, Z. (2011) RegulaDon of axonal mitochondria trasnport and its impact on synapDc transmission. Neuroscience Research. 70(1): 9-­‐15. Myo5a in neurons and mice Cai, Q. Davis, ML. Sheng, Z. (2011) RegulaDon of axonal mitochondria trasnport and its impact on synapDc transmission. Neuroscience Research. 70(1): 9-­‐15. Normal Myo5a-­‐ h=p://jaxmice.jax.org/jaxnotes/512/512l.html h=ps://database.riken.jp/sw/en/B10.D1-­‐Myo5a_d__Hir References • 
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h=p://www.cellmigraDon.org/resource/discovery/vale h=p://www.equinenow.com/farm/cressant_hill_arabians.htm h=p://arabiansltd.com/recent-­‐arabian-­‐sales h=p://delightarabhorsestud.homestead.com/Foals_.html h=p://animalgeneDcs.eu/Equine/equine-­‐geneDc-­‐disease/lfs.html h=p://livingmydreamkn.blogspot.com/2011/04 Divers, TJ. (2006) Structural and FuncDonal Diseases of the Equine Brain. Clinical Techniques in Equine prac1ce. 5(1): 67-­‐73 h=p://www.more4kids.info/699/childrens-­‐intuiDon/ h=p://www.fcarreras.org/en/mohammed-­‐griscelli-­‐syndrome-­‐former-­‐paDent_42620 h=p://jaxmice.jax.org/jaxnotes/512/512l.html h=ps://database.riken.jp/sw/en/B10.D1-­‐Myo5a_d__Hir Kneussel, M. Wagner, W. (2013) Myosin motors and neuronal synapses: drivers of membrane transport and acDn dynamics. Nature Reviews: Neuroscience. 14: 233-­‐48. Clustal Omega h=p://www.ebi.ac.uk/Tools/msa/clustalo/ Huber, L. A. (2003) Is proteomics heading in the wrong direcDon? Nature Reviews Molecular Cell Biology, 4, 74-­‐80 h=p://kaDya-­‐junkyard.blogspot.com/2013/03/physiology-­‐of-­‐color-­‐and-­‐pa=ern-­‐in.html Kuroda, T.S. et al. (2003). The acDn binding domain of slac-­‐2a/melanophilin is required for melanosome distribuDon in melanocytes. Mol Cel Biol. 23(15): 5245-­‐55. h=p://post.jagran.com/sakhi-­‐Zebrafish-­‐to-­‐mend-­‐a-­‐broken-­‐heart-­‐1321348010 h=p://www.erasmusmc.nl/klinische_geneDca/research/intro/willemsen/zebrafish/?lang=en Luo, N. et al. (2012) Evidence for a role of inositol polyphosphate 5-­‐phosphatase INPP5E in cilia formaDon in zebrafish. Vision Research. 75:98-­‐107. String h=p://string-­‐db.org/ Marks, MS. Seabra, MC. (2001) The Melanosome: Membrane Dynamics in Black and White. Nature Reviews: Molecular Cell Biology. 2:1-­‐11. Cai, Q. Davis, ML. Sheng, Z. (2011) RegulaDon of axonal mitochondria trasnport and its impact on synapDc transmission. Neuroscience Research. 70(1): 9-­‐15. QuesDons? h=p://www.cellmigraDon.org/resource/discovery/vale