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
h=p://livingmydreamkn.blogspot.com/2011/04

LFS
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 Myo5a-

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

Normal

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 Myo5a-

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

Normal

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 unidenDed 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 sh to study Melanosomes?

h=p://post.jagran.com/sakhi-Zebrash-to-mend-a-broken-heart-1321348010

h=p://www.erasmusmc.nl/klinische_geneDca/research/intro/willemsen/ zebrash/?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 zebrash. Vision Research. 75:98-107.

Sort proteins using gene ontology

Vesicle Transport

Zebra sh 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 Expected

Luo, N. et al. (2012) Evidence for a role of inositol polyphosphate 5- phosphatase INPP5E in cilia formaDon in zebrash. Vision Research. 75:98-107.

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
h=p://jaxmice.jax.org/jaxnotes/512/512l.html

Myo5a-
h=ps://database.riken.jp/sw/en/B10.D1-Myo5a_d__Hir

References
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-Zebrash-to-mend-a-broken-heart-1321348010 h=p://www.erasmusmc.nl/klinische_geneDca/research/intro/willemsen/zebrash/?lang=en Luo, N. et al. (2012) Evidence for a role of inositol polyphosphate 5-phosphatase INPP5E in cilia formaDon in zebrash. 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