F44E5'4 Heat Shock Protein 70

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F44E5.4 (Heat Shock Protein 70)
IMBBR 315
Christopher Ricupero
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Clones
58.3 of colonies picked vs. successful clones
sequenced
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Clones
58.3 of colonies picked vs. successful clones
sequenced
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14D6-57 Sanger BLAST N Results

• Insert size small 418 bp
• Sequenced 1 direction
• Spanned entire insert

Exon
Intron
Exon
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14D6-57 Sanger BLAST N Results

• Insert size small 418 bp
• Sequenced 1 direction
• Spanned entire insert

• Interesting Observation
• Sequence matching two unique genomic regions, in
  two directions

What is going on?
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Sequence Summary for F44E5

• P(N) 8.6 e-65 - Sequence matches hsp70 (Heat
  Shock Protein)
• Gene duplication
• Protein - 645 AAs
• RNAi - Wild type morphology

What is this protein?
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Structure

• Size 70kD
• Length 645 AAs
• ATP dependent
• Single Monomer
• 3 Domains
• N Terminal - ATP Binding and hydrolysis (44 kDa)
  Homologous to Actin and Hexokinase
• Substrate Binding Hydrophobic regions (15-20
  kDa)
• C-terminal Lid for substrate binding (15 -20
  kDa)
• Allosteric properties conformational changes

What does it do?
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Hsp70 (Heat Shock Protein)
Essential in Protein Folding
Cellular stress protection

• Key Points
• A Molecular chaperone
• Inducible during stressful conditions (heat,
  toxins, etc..)
• Ubiquitous expression - In all cells at all
  biological levels
• Highly conserved family (Hsp 40, 60, 90 etc..)
• Consists of stress-inducible and constitutive
  family members (Hsc 70, Hsp72)

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Hsp70 - Molecular chaperone

• Recognizes binds regions rich in hydrophobic
  residues
• Stabilizes prevents misfolding by open
  conformation
• Prevents aggregation and degradation during
  synthesis
• Multiple Hsp70 complexes per polypeptide chain
• Releases or transfers to other chaperones/chaperon
  ins (Hsp60, Hsp90)

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Hsp70 Cellular Protection

• Stress Induced Prevents protein denaturation
  and incorrect polypeptide aggregation during
  exposure to physiochemical insults
• Hsp70 expression linked in several different
  models of neurodegenerative diseases

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Hsp70 Neurodegenerative Diseases
Hsp 70
Fonte, Virginia et al. (2002) Proc. Natl. Acad.
Sci. USA 99, 9439-9444

• C. Elegans model of Alzheimers disease
• Hsp70 and Hsp16 expression patterns correlated
  with phenotype

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Hsp70 - Additional Functions
Involved in various cellular functions and
diseases
Apoptosis
Clathrin cage disassembly
Parkinson's Disease
Dauer Stage
Translocation
Cancer
Prions
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Acknowledgements
Thank you! Dr. Marty Nemeroff Dr. Monica
Driscoll Indrani Chatterjee Brian Gelfand
The rest of 315 staff
Questions?