Amyotrophic Lateral Sclerosis and SOD 1 Anne Sapiro

Amyotrophic Lateral Sclerosis and SOD 1 Anne Sapiro

Amyotrophic lateral sclerosis facts Neurodegenerative disease Loss of motor control Fatal within 3 -5 years 90% sporadic 20% of familial ALS caused by mutations in SOD 1 www. thehealthsuccesssite. com

Superoxide Dismutase 1 (SOD 1) Furukawa Y , O'Halloran T V J. Biol. Chem. 2005; 280: 17266 -17274

ALS-causing mutations in SOD 1 Human SOD 1 1 5 150 154 aa SOD Cu 154 5 83 amino acids with missense or nonsense mutations http: //alsod. iop. kcl. ac. uk

SOD 1 protein domains across homologs Human SOD 1 1 5 Mouse SOD 1 1 5 Zebrafish SOD 1 150 154 aa SOD Cu

Mutant SOD 1: Toxic GOF Furukawa Y , O'Halloran T V J. Biol. Chem. 2005; 280: 17266 -17274

Gene Ontology: Biological Processes SOD 1 Apoptosis Mitochondrial Respiration

SOD 1 aggregates lead to mitochondrial defects What comprises aggregates?

Human SOD 1 Interaction Network

Aggregates and neurodegeneration Parkinson’s Lewy bodies Huntington’s inclusions http: //neuropathology-web. org/ Alzheimer’s plaques http: //missinglink. ucsf. edu/lm/ids_104_neurodegenerative/Case 2 Micro. htm http: //cdn. physorg. com

Parkinson’s Lewy bodies α-synuclein Neurofilaments Adapted from Branco et al. 2010 Ubiquitin

Hypothesis: ALS aggregates contain components of Parkinson’s Lewy bodies. Approach: Look at interaction networks in humans and homologs

Ubiquitin-related proteins in SOD 1 interaction networks Uba 52 RNF 19 A E 3 Ubiquitin-protein ligase

Neurofilaments in SOD 1 interaction networks

Question: What else is in ALS aggregates? Approach: Proteomics in model organism

SOD 1 phylogeny ✔ ✔

Zebrafish SOD 1 interaction network

Zebrafish SOD 1 and PARK 7 interaction network

SOD 1 and PARK 7 in mitochondria

Gene Ontology: Biological Processes SOD 1 Apoptosis Mitochondrial Respiration

Mitochondrial respiration http: //pages. slu. edu/faculty/kennellj/images/respchain 2. gif

Mitochondrial respiration http: //pages. slu. edu/faculty/kennellj/images/respchain 2. gif

High incidence of mitochondrial proteins in interaction networks

Question: What else is in ALS aggregates? Hypothesis: ALS aggregates will include SOD 1, Ubiquitin, Neurofilaments, mitochondrial proteins. Approach: Proteomic analysis of aggregates in SOD 1 mutant zebrafish

Experimental Design G 93 R-SOD 1 overexpression mutant Culture motor neurons Mud. PIT on aggregates to identify components Subcellular fractionation

Hypothetical results

SOD 1 aggregates lead to mitochondrial defects

Hypothesis: Mutations in genes encoding mitochondrial proteins in aggregates cause ALS.

Genes known to cause ALS CDH 22 ALS 2 ANG CHMP 2 B ATXN 2 CNTN 6 BCL 11 B CRIM 1 CRYM BCL 6 C 9 orf 72 DAO CDH 13 DCTN 1 DIAPH 3 DOC 2 B OMA 1 SQSTM 1 EWS OPTN SYT 9 FEZF 2 PCP 4 TAF 15 FIG 4 RAMP 3 TARDBP GRB 14 SETX UBQLN 2 LUM SIGMAR 1 VAPB NEFH SOX 5 VCP NETO 1 SPG 11 http: //alsod. iop. kcl. ac. uk/misc/data. Download. aspx#C 1

Genes known to cause ALS CDH 22 DOC 2 B OMA 1 SQSTM 1 ALS 2 EWS OPTN SYT 9 ANG CHMP 2 B FEZF 2 PCP 4 TAF 15 ATXN 2 CNTN 6 Mitochondrial Metalloendopeptidase OMA 1 TARDBP CRIM 1 FIG 4 RAMP 3 BCL 11 B Negative regulator CRYM GRB 14 of SETX UBQLN 2 BCL 6 mitochondrial fusion LUM SIGMAR 1 VAPB C 9 orf 72 DAO NEFH SOX 5 VCP CDH 13 DCTN 1 DIAPH 3 NETO 1 SPG 11 http: //alsod. iop. kcl. ac. uk/misc/data. Download. aspx#C 1

Zebrafish SOD 1 and PARK 7 interaction network

Proposed model m. SOD 1 Disturbance of mitochondrial respiration; apoptosis, etc. Abnormal mitochondrial dynamics

Future experiments Further characterization Chemical genetics: of OMA 1 mitochondrial proteins in fusion and fission (Tateno et al. 2009)

Questions?