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Inhibition of HIV‐1 transcription by a tunable chimeric tRNA(Ser)‐nucloelar localizing trans‐activation response element (TAR) decoy
Author(s) -
Stevens Christian,
Scherer Lisa,
Rossi John,
Haushalter Karl
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.985.1
Subject(s) - decoy , transcription (linguistics) , rna , microbiology and biotechnology , chemistry , activator (genetics) , transfer rna , biology , messenger rna , luciferase , transfection , biochemistry , gene , receptor , philosophy , linguistics
In order for HIV‐1 to achieve high‐level transcription of its integrated proviral DNA, the trans‐activator of transcription (Tat) protein must bind to the TAR region of the nascent viral RNA transcript. A TAR decoy is a short RNA oligomer that inhibits HIV‐1 transcription by binding to the Tat protein, thereby competitively inhibiting the TAR‐tat interaction. Previous work has shown that nucleolar‐localization of a TAR decoy by fusion with the U16 small nucleolar RNA (snoRNA) backbone dramatically increases the TAR decoy inhibition efficiency. In this work, the nucleolar‐localized TAR decoy is expressed as the 3′ trailing sequence of a tRNA(Ser) chimeric transcript. Maturation of the tRNA chimera by tRNA processing enzymes releases the U16‐TAR element from the tRNA. Using a dual‐luciferase reporter assay with firefly luciferase under the control of the HIV‐1 long terminal repeat (LTR) promoter, we show that expression of the tRNA‐TAR decoy chimera inhibits HIV‐1 transcription in human HCT116 cells. We also show that mutations in the tRNA(Ser) acceptor stem allow for inhibition efficiency to be tunable.