U3
Download
1 / 1

Comparative Transcription Analysis of - PowerPoint PPT Presentation


  • 50 Views
  • Uploaded on

U3. U5. R. c-Ets-1 ccAGGAAgttaataa. CDP ccataCTTCTTGCTG. HIF-1 agtACGTGcctc. STAT5A ctctaggcacgaaaCTTCTtggaa. STAT5A aacttcttGGAAAct. GATA-4 AGATAacaggga. IRF gaaagAGAAAa. AREB6. Comparative Transcription Analysis of

loader
I am the owner, or an agent authorized to act on behalf of the owner, of the copyrighted work described.
capcha
Download Presentation

PowerPoint Slideshow about ' Comparative Transcription Analysis of ' - tad


An Image/Link below is provided (as is) to download presentation

Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author.While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server.


- - - - - - - - - - - - - - - - - - - - - - - - - - E N D - - - - - - - - - - - - - - - - - - - - - - - - - -
Presentation Transcript

U3

U5

R

c-Ets-1

ccAGGAAgttaataa

CDP

ccataCTTCTTGCTG

HIF-1

agtACGTGcctc

STAT5A

ctctaggcacgaaaCTTCTtggaa

STAT5A

aacttcttGGAAAct

GATA-4

AGATAacaggga

IRF

gaaagAGAAAa

AREB6

Comparative Transcription Analysis of

Different PERV LTR Subtypes in Human, Monkey and Pig cell lines

Yi-Deun Jung1, Jae-Won Huh1,2, Dae-Soo Kim3, Hong-Seok Ha1 and Heui-Soo Kim1

1 Division of Biological Sciences, College of Natural Sciences, Pusan National University, Busan 609-735, Republic of Korea

2 National Primate Research Center, KRIBB, Ochang, Chungbuk 363-883, Republic of Korea

3 Korea Bioinformation Center, KRIBB, Daejeon 305-806, Republic of Korea

http://www.primate.or.kr

  • Abstract

Porcine endogenous retroviruses (PERV) in the pig genome have the potential to act as harmful factors in xenotransplantation (pig-to-human). Long terminal repeats (LTRs) of the porcine endogenous retroviruse (PERV) elements show promoter activity that can affect human functional genes. Transcription activity of 11 different long terminal repeat (LTR) subtypes of porcine endogenous retroviruse (PERV) are analyzed using human (HEK293), monkey (Cos7), and pig(PK15) kidney cell line. They show the varied transcription regulation activity in different cell lines. However, E4 type (H2-2) shows the distinct promoter activity in the human kidney HEK293 cell line. We thus constructed the series of deletion mutants based on the boundary of U3, R and U5 region. Transient transfection assay of 10 deletion mutants show that U3 is crucial region for transcription regulation. By the computational approaches, 7 potential transcription factors are identified in U3 region. MAZR, HIF-1, STAT5A, and GATA-4 seem to be positive factors and c-ETS-1, CDP, and IRF should be acted by negative elements in human cell lines.

  • Introduction

  • Materials & Methods

Virus (Virus-like particle)

Virus

11. Budding

1.Receptor

interaction

10. Virus assembly

(Virus –like particles)

9. Translation

Proteins

RNA

Sample

RT-PCR

cloning

Sequencing

&

analysis

Cap

(A)n

(A)n

Cap

Viral genomic RNA

Viral spliced RNA

4. Reverse

Transcription

8. RNA transport

7. RNA processing

6.Transcription

3. Uncoating

U3

U5

R

gag

pol

env

LTR

LTR

Recombination

5. Integration

2. Penetration

Luciferase

assay

Deletion

mapping

Subtype

identification

Solitary LTR

gag : coding for matrix and capsid proteins

pol : reverse transcriptase and integrase proteins

env : product viral envelope and transmembrane structures

U3

R

U5

U3

R

U5

gag

pol

env

U3

R

U5

PBS

3’UTR

5’UTR

  • Results & Discussion

  • Deletion mutant structure

  • Transient transfection assay of PERV-LTR in different cell lines

1

2

3

4

5

6

7

8

9

10

  • Luciferase reporter gene assay for 10 deletion mutant

1) Separated by structure

2) Separated by transcription factor (U3 region)

  • PERV LTR structure

TATA

U3

R

U5

3) Separated by transcription factor

( deletion mutant 3 )

606 bp

67 bp

26 bp

  • Reference

  • Patience C, Takeuchi Y, Weiss RA (1997) Infection of human cells by an endogenous retrovirus of pigs. Nat Med 3: 276-282.

  • Scheef G, Fischer N, Krach U, Tonjes RR (2001) The number of a U3 repeat box acting as an enhancer in long terminal repeats of polytropic replication-competent porcine endogenous retroviruses dynamically fluctuates during serial virus passages in human cells. J Virol 75: 6933-6940


ad