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Who is the Solitary Long Terminal Repeats of ERV-9 Endogenous Retrovirus?

Who is the Solitary Long Terminal Repeats of ERV-9 Endogenous Retrovirus?. 석사 1 년 김 우 연. Contents. Identification of ERV9 LTR Identification of R within ERV9 LTR Mutational Analysis of ERV9 Promoter Region Mobilization of ERV9 during Primate Evolution

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Who is the Solitary Long Terminal Repeats of ERV-9 Endogenous Retrovirus?

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  1. Who is the Solitary Long Terminal Repeats of ERV-9 Endogenous Retrovirus? 석사 1 년 김 우 연

  2. Contents • Identification of ERV9 LTR • Identification of R within ERV9 LTR • Mutational Analysis of ERV9 Promoter Region • Mobilization of ERV9 during Primate Evolution • ERV9 LTR in β-globin gene and Axin gene • Evolution of ERV9 • Future study

  3. 1. Identification of ERV9 in undifferentiated embryonal carcinoma cells

  4. Assay of ERV9 LTR promoter activity in different human cell lines

  5. 2. Identification of R within ERV9 LTR Identification of R within HERV9 LTR ( LTR12 )

  6. Identification of transcription start sites - Transfection in NT2/D1 cells - Total RNA isolation - Primer extension analysis Using a primer complementary to the CAT coding region

  7. CAT expression assay • CAT gene • Chloramphenicol Acetyltransferase • counting the amount of [14C] Chloramphenicol converted to the mono-acetylated forms • Efficient promoter activity region : -70/+6, -70/-7

  8. DNase I footprint analysis : +7 ~ -10 : -33 ~ -54 Sp1 transcription factor binds to -54 ~ -33

  9. Sp1 transcription factor binds to -54 ~ -33

  10. 3. Mutational Analysis of ERV9 Promoter Region

  11. The potential of different transcription factors

  12. 4. Mobilization of ERV9 during Primate Evolution

  13. zinc finger protein 80 (ZNF80) gene

  14. ZNF80 in the primate species

  15. Alignment of LTRs regions

  16. Phylogenetic tree of radiation, insertion time Insertion time of the ERV9 element

  17. 5. ERV9 LTR in β-globin gene and Axin gene ERV9 LTR is conserved during primate evolution and possess enhancer activities

  18. β-globin gene region • A. • nuclease hypersensitive site 5,4,3,2,1 • HBE1, HBG2,1 HBBP1, HBD, HBB • B. • Human, Chimpanzee, Gorilla, Orangutan – LTR12 • Gibbon, Monkey – no LTR12

  19. Axin gene - HESAS Result of HESAS

  20. Axin gene – axin LTR

  21. By FACS

  22. Enhancer and promoter activities

  23. 6. Evolution of HERV9

  24. Identified ERV9 Elements Synapomorphic combination of character states i.e. Diagnostic nucleotide substritutions

  25. Ages of ERV9 Subfamilies

  26. Phylogenetic relationships

  27. Future study • ERV9 LTR ( LTR11, LTR12, LTR17, pTR5 ) 분석 • U5 영역의 repeat sequence 분석 • Human genome 전체에서 ERV9 LTR 삽입 상태 분석 • Gene 과 연결된 ERV9 LTR 분석 • Gene 과 연결된 상태에 따라 분석 • ERV9 LTR의 Expression 분석

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