Construction of a reporter vector for analysis of bidirectional transcriptional activity of retrovirus LTR.

Abstract:

:To study the transcriptional activity of the HIV-1 LTR, we constructed a vector containing Renilla and Firefly luciferase genes under the control of the LTR (wild-type or mutated version) and oriented in a manner that allowed them to be transcribed in opposite directions. We found that the HIV-1 LTR acted as a bidirectional promoter, which activity was controlled by NF-κB- and Sp1-binding sites in both orientations. We next analyzed with this reporter vector the bidirectional promoter activity of the HTLV-1 LTR and showed that this LTR also possessed a bidirectional transcriptional activity. Interestingly, Sp1-binding elements were also involved in the control of HTLV-1 bidirectional transcription. Moreover, both retroviral trans-activators, Tat and Tax, could preferentially activate sense transcription with no or limited effect on the extent of antisense transcription. We also cloned into this plasmid the MLV LTR and found that the LTR of a simple retrovirus also possessed bidirectional transcriptional activity. This reporter vector represents a powerful tool to analyze the bidirectional transcriptional activity of retrovirus LTRs.

journal_name

Plasmid

journal_title

Plasmid

authors

Arpin-André C,Laverdure S,Barbeau B,Gross A,Mesnard JM

doi

10.1016/j.plasmid.2014.06.001

subject

Has Abstract

pub_date

2014-07-01 00:00:00

pages

45-51

eissn

0147-619X

issn

1095-9890

pii

S0147-619X(14)00038-9

journal_volume

74

pub_type

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