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DNA Methylation Patterns & Epigenetic Regulation in the Pacific Oyster  Mackenzie Gavery & Steven Roberts University of Washington School of Aquatic and Fishery Sciences
Outline ,[object Object]
Results: characterization of DNA                          methylation in Pacific oysters
Implications
Future directions,[object Object]
Background:  color disease resistance growth TRAITS pathogens toxins nutrition EPIGENOME (DNA methylation) ENVIRONMENT GENES (DNA)
Background:  color disease resistance growth TRAITS pathogens toxins nutrition EPIGENOME (DNA methylation) ENVIRONMENT GENES (DNA)
Epigenetics  Heritable changes in trait or phenotype, caused by a mechanism other than mutation to the DNA sequence Most well understood epigenetic mechanism is DNA methylation: ,[object Object]
regulates gene expression
influenced by the environment Me C G G C
DNA methylation Effects of disruptions: tumor promotion alteration of development inhibition of reproduction Compounds that impact normal epigenetic functions: Endocrine disruptors: estrogen, BPA, pesticides
Characterization of DNA methylation in Pacific oysters: describe distribution of methylation elucidate functional significance
Results Methylation Specific PCR Bisulfite sequencing In silicoanalysis
Results Methylation Sensitive PCR Bisulfite sequencing In silicoanalysis
Results: gene-targeted approach Methylation Sensitive PCR 5 stress related genes were examined Identified CpG methylation in heat shock protein 70 Bisulfite sequencing 136 bp fragment: 1 of 7 CpG methylated               (homology to neuromedin-u receptor) 93 bp fragment: 1 of 2 CpG methylated                 (homology to bromodomain adjacent to zinc finger domain)
Results Methylation Sensitive PCR Bisulfite sequencing In silicoanalysis
Results Methylation Sensitive PCR Bisulfite sequencing In silicoanalysis predicted methylation status of 12,000 C. gigas sequences from GigasBase sequences were grouped by Gene Ontology term an average predicted methylation status was determined
Regulation of Gene Expression low mid high Predicted DNA Methylation
Regulation of Gene Expression low mid high Predicted DNA Methylation
Implications: evidence suggests DNA methylation plays a regulatory role in Pacific oysters implications for immune/stress responses
Implications: Environment   LowHigh
Implications: Selective Breeding Selective breeding can contribute to improved & predictable performance in oysters Understanding geneticand epigenetic influences will increase predictability
Implications: Hybrid Vigor Heterosis (hybrid vigor) mechanism not fully understood epigenetic mechanisms have been proposed better understanding will allow for greater control in predicting and manipulating gene expression in oysters  X =

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NSA Feb 2010: DNA Methylation Patterns & Epigenetic Regulation in the Pacific Oyster

  • 1. DNA Methylation Patterns & Epigenetic Regulation in the Pacific Oyster Mackenzie Gavery & Steven Roberts University of Washington School of Aquatic and Fishery Sciences
  • 2.
  • 3. Results: characterization of DNA methylation in Pacific oysters
  • 5.
  • 6. Background: color disease resistance growth TRAITS pathogens toxins nutrition EPIGENOME (DNA methylation) ENVIRONMENT GENES (DNA)
  • 7. Background: color disease resistance growth TRAITS pathogens toxins nutrition EPIGENOME (DNA methylation) ENVIRONMENT GENES (DNA)
  • 8.
  • 10. influenced by the environment Me C G G C
  • 11. DNA methylation Effects of disruptions: tumor promotion alteration of development inhibition of reproduction Compounds that impact normal epigenetic functions: Endocrine disruptors: estrogen, BPA, pesticides
  • 12. Characterization of DNA methylation in Pacific oysters: describe distribution of methylation elucidate functional significance
  • 13. Results Methylation Specific PCR Bisulfite sequencing In silicoanalysis
  • 14. Results Methylation Sensitive PCR Bisulfite sequencing In silicoanalysis
  • 15. Results: gene-targeted approach Methylation Sensitive PCR 5 stress related genes were examined Identified CpG methylation in heat shock protein 70 Bisulfite sequencing 136 bp fragment: 1 of 7 CpG methylated (homology to neuromedin-u receptor) 93 bp fragment: 1 of 2 CpG methylated (homology to bromodomain adjacent to zinc finger domain)
  • 16. Results Methylation Sensitive PCR Bisulfite sequencing In silicoanalysis
  • 17. Results Methylation Sensitive PCR Bisulfite sequencing In silicoanalysis predicted methylation status of 12,000 C. gigas sequences from GigasBase sequences were grouped by Gene Ontology term an average predicted methylation status was determined
  • 18. Regulation of Gene Expression low mid high Predicted DNA Methylation
  • 19. Regulation of Gene Expression low mid high Predicted DNA Methylation
  • 20. Implications: evidence suggests DNA methylation plays a regulatory role in Pacific oysters implications for immune/stress responses
  • 22. Implications: Selective Breeding Selective breeding can contribute to improved & predictable performance in oysters Understanding geneticand epigenetic influences will increase predictability
  • 23. Implications: Hybrid Vigor Heterosis (hybrid vigor) mechanism not fully understood epigenetic mechanisms have been proposed better understanding will allow for greater control in predicting and manipulating gene expression in oysters X =
  • 24.
  • 25. Future Directions Method evaluation/development: challenges associated with non-model species new approaches: Whole genome bisulfite sequencing (BS-seq) Methylated DNA immunoprecipitation (MeDIP) MeDIP-seq MeDIP-chip
  • 26. Summary Characterization of DNA methylation in Pacific oyster suggests a role in gene regulation, specifically genes with inducible expression DNA methylation could be an important mechanism contributing to phenotypic variation in oysters Important evaluate & develop methods and tools to evaluate epigenetic mechanisms in bivalves
  • 27. Acknowledgements UW, SAFS Dr. Steven Roberts Samuel White Lisa Crosson Emma Timmins-Schiffman Taylor Shellfish Farms Joth Davis NSA-PCS NOAA Aquaculture Program