Pusan National UniversityInterdisciplinary Program of Bioinformatics
Who is the Solitary Long Terminal Who is the Solitary Long Terminal Repeats of ERV-9 Endogenous Repeats of ERV-9 Endogenous Retrovirus?Retrovirus?
석사 1 년김 우 연
ContentsContents
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
1. Identification of ERV9 in 1. Identification of ERV9 in undifferentiated embryonal carcinoma undifferentiated embryonal carcinoma cellscells
Assay of ERV9 LTR promoter activity in Assay of ERV9 LTR promoter activity in different human cell linesdifferent human cell lines
2. Identification of R within ERV9 2. Identification of R within ERV9 LTRLTR
Identification of R within HERV9 LTR ( LTR12 )
Identification of transcription Identification of transcription start sitesstart sites
- Transfection in NT2/D1 cells- Total RNA isolation- Primer extension analysis Using a primer complementary to the CAT coding region
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
DNase I footprint analysis: +7 ~ -10: -33 ~ -54Sp1 transcription factor binds to-54 ~ -33
Sp1 transcription factor binds to-54 ~ -33
3. Mutational Analysis of ERV9 3. Mutational Analysis of ERV9 Promoter RegionPromoter Region
The potential of different transcription The potential of different transcription factorsfactors
4. Mobilization of ERV9 during Primate 4. Mobilization of ERV9 during Primate EvolutionEvolution
zinc finger protein 80 (ZNF80) zinc finger protein 80 (ZNF80) genegene
ZNF80 in the primate speciesZNF80 in the primate species
Alignment of LTRs regionsAlignment of LTRs regions
Phylogenetic tree of radiation, Phylogenetic tree of radiation, insertion timeinsertion time
Insertion time of the ERV9 element
5. ERV9 LTR in 5. ERV9 LTR in β -globin gene and Axin gene
ERV9 LTR is conserved during primate evolution and possess enhancer activities
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
Axin gene - HESASAxin gene - HESAS
Result of HESAS
Axin gene – axin LTRAxin gene – axin LTR
By FACS
Enhancer and promoter activitiesEnhancer and promoter activities
6. Evolution of HERV96. Evolution of HERV9
Synapomorphic combination of character statesi.e.Diagnostic nucleotide substritutions
Identified ERV9 ElementsIdentified ERV9 Elements
Ages of ERV9 SubfamiliesAges of ERV9 Subfamilies
Phylogenetic relationshipsPhylogenetic relationships
Future studyFuture 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 LTR 의 Expression 분석