INT303399

Context Info Confidence 0.50 First Reported 2010 Last Reported 2010 Negated 0 Speculated 0 Reported most in Body Documents 1 Total Number 11 Disease Relevance 0.85 Pain Relevance 0

plasma membrane (Jag1) cytoplasm (Myh3)

Anatomy Link Frequency myoblast 3 thick filaments 2 muscle 1 tail region 1 head 1 Jag1 (Mus musculus) Myh3 (Mus musculus)

Pain Link Frequency Relevance Heat Spinal cord 11 47.96 cytokine 11 19.28 Sciatic nerve 11 19.12 Pain 11 15.60 imagery 22 5.00

Disease Link Frequency Relevance Heat Frailty 44 95.16 Hypertrophy 11 91.64 Muscular Atrophy 99 90.20 Injury 11 66.04 Nutritional Deficiencies 11 54.64 Pain 11 15.60 Sprains And Strains 11 5.00 Targeted Disruption 11 5.00 Stress 11 5.00 Appetite Loss 11 5.00

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Key:

Protein

Mutation

Event

Anatomy

Negation

Speculation

Pain term

Disease term

The fact that Ozz and its direct partner Elo C could be co-immunoprecipitated from sarcomeric but not soluble myosin extracts of E16.5 embryonic muscle indicates that at this age a proportion of Ozz molecules bound to MyHCemb is assembled into the E3 ligase complex.

Ozz Binding (bound) of MyHCemb in muscle

1) Confidence 0.50 Published 2010 Journal PLoS ONE Section Body Doc Link PMC2844429 Disease Relevance 0.05 Pain Relevance 0

We found that Ozz recognizes the rod portion of MyHCemb, which forms the core of sarcomeric thick filaments where it is not easily accessed for binding or exchange by soluble cytoplasmic molecules.

Ozz Binding (recognizes) of MyHCemb in thick filaments

2) Confidence 0.50 Published 2010 Journal PLoS ONE Section Body Doc Link PMC2844429 Disease Relevance 0 Pain Relevance 0

Direct interaction of Ozz with myofilamentous MyHCemb was proven by co-immunoprecipitation of Ozz with anti-MyHCemb antibody only from the insoluble preparations (P), but not from the soluble preparations (S), albeit the amount of Ozz was greater in the latter (Fig. 5B, panels 2 and 6).

Ozz Binding (interaction) of MyHCemb

3) Confidence 0.44 Published 2010 Journal PLoS ONE Section Body Doc Link PMC2844429 Disease Relevance 0 Pain Relevance 0

Ozz-MyHCemb complexes were detected in crude lysates of myoblast by immunoprecipitation with anti-MyHCemb antibody, followed by Western blotting of the immunoprecipitates with the anti-Ozz antibody.

Ozz Binding (complexes) of MyHCemb in myoblast

4) Confidence 0.39 Published 2010 Journal PLoS ONE Section Body Doc Link PMC2844429 Disease Relevance 0 Pain Relevance 0

Here we present evidence that the Ozz-E3 ligase, by binding to the rod domain of a fully assembled MyHCemb, marks it for ubiquitination and degradation, probably facilitating the subsequent assembly of new isoforms.

Ozz Binding (binding) of MyHCemb

5) Confidence 0.37 Published 2010 Journal PLoS ONE Section Body Doc Link PMC2844429 Disease Relevance 0.38 Pain Relevance 0

We found that Ozz recognizes the rod portion of MyHCemb, which forms the core of sarcomeric thick filaments where it is not easily accessed for binding or exchange by soluble cytoplasmic molecules.

Ozz Binding (recognizes) of MyHCemb in thick filaments

6) Confidence 0.37 Published 2010 Journal PLoS ONE Section Body Doc Link PMC2844429 Disease Relevance 0 Pain Relevance 0

However, under these stringent experimental conditions (high salt concentration), while we were able to demonstrate Ozz binding with its direct interacting partners, MyHCemb and Elo C, we could not co-immunoprecipitate the remaining components of the Ozz-E3 complex (not shown).

Ozz Binding (binding) of MyHCemb

7) Confidence 0.34 Published 2010 Journal PLoS ONE Section Body Doc Link PMC2844429 Disease Relevance 0 Pain Relevance 0

Ozz binding to MyHCemb differs from that of other E3 ligases in that it targets the tail portion of assembled sarcomeric myosin rather than the head portion of soluble myosin, and we suggest that these properties are fundamental to its role in muscle development as opposed to muscle atrophy.

Ozz Binding (binding) of MyHCemb in head associated with muscular atrophy

8) Confidence 0.33 Published 2010 Journal PLoS ONE Section Body Doc Link PMC2844429 Disease Relevance 0.23 Pain Relevance 0

To verify whether the Ozz-MyHCemb interaction occurred in vivo, crude lysates of proliferating (day 0), differentiating (day 2) and terminally differentiated (day 4) primary myoblast cultures prepared from newborn wild-type mice were immunoprecipitated with anti-MyHCemb antibody or an isotype matching control IgG, and probed on immunoblots with anti-Ozz antibody.

Ozz Binding (interaction) of MyHCemb in myoblast

9) Confidence 0.33 Published 2010 Journal PLoS ONE Section Body Doc Link PMC2844429 Disease Relevance 0 Pain Relevance 0

To confirm Ozz' interaction with the tail of MyHCemb and to identify the minimal regions of the tail needed for this interaction, we performed a series of 2-hybrid experiments using as preys either the full length tail domain (residues 1040–1941), the full length head/neck domain (residues 1–1040), or several deletion mutants of the tail region (Fig. 1E and Fig.

Ozz Binding (interaction) of MyHCemb in tail region

10) Confidence 0.33 Published 2010 Journal PLoS ONE Section Body Doc Link PMC2844429 Disease Relevance 0.19 Pain Relevance 0

To verify whether the Ozz-MyHCemb interaction occurred in vivo, crude lysates of proliferating (day 0), differentiating (day 2) and terminally differentiated (day 4) primary myoblast cultures prepared from newborn wild-type mice were immunoprecipitated with anti-MyHCemb antibody or an isotype matching control IgG, and probed on immunoblots with anti-Ozz antibody.

Ozz Binding (interaction) of MyHCemb in myoblast

11) Confidence 0.33 Published 2010 Journal PLoS ONE Section Body Doc Link PMC2844429 Disease Relevance 0 Pain Relevance 0

General Comments

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