Use of a Polyethylene Glycol–Peptide Conjugate in a Competition Gel Shift Assay for Screening Potential Antagonists of HIV-1 Tat Protein Binding to TAR RNA

1995 ◽  
Vol 232 (2) ◽  
pp. 238-242 ◽  
Author(s):  
Jihong Wang ◽  
Shaei-Yun Huang ◽  
Indrani Choudhury ◽  
Michael J. Leibowitz ◽  
Stanley Stein
Science ◽  
1990 ◽  
Vol 249 (4974) ◽  
pp. 1281-1285 ◽  
Author(s):  
K. Weeks ◽  
C Ampe ◽  
S. Schultz ◽  
T. Steitz ◽  
D. Crothers
Keyword(s):  
Tar Rna ◽  

1998 ◽  
Vol 26 (12) ◽  
pp. 2886-2890 ◽  
Author(s):  
A. Garbesi ◽  
F. Hamy ◽  
M. Maffini ◽  
G. Albrecht ◽  
T. Klimkait
Keyword(s):  
Tar Rna ◽  

Virology ◽  
1991 ◽  
Vol 182 (2) ◽  
pp. 570-577 ◽  
Author(s):  
James Kamine ◽  
Paul Loewenstein ◽  
Maurice Green
Keyword(s):  
Tar Rna ◽  

1993 ◽  
Vol 21 (1) ◽  
pp. 151-154 ◽  
Author(s):  
Richard W. Barnett ◽  
Ulrike Delling ◽  
Raya Kuperman ◽  
Nahum Sonenberg ◽  
Martin Sumner-Smith

2009 ◽  
Vol 106 (29) ◽  
pp. 11931-11936 ◽  
Author(s):  
A. Davidson ◽  
T. C. Leeper ◽  
Z. Athanassiou ◽  
K. Patora-Komisarska ◽  
J. Karn ◽  
...  

2003 ◽  
Vol 84 (3) ◽  
pp. 603-606 ◽  
Author(s):  
Lars H. Lund ◽  
Britta Wahren ◽  
Mariano A. Garcia-Blanco

Human immunodeficiency virus type 1 (HIV-1) Tat and human Cyclin T1 form a complex and together recognize the viral TAR RNA element with specificity. Using HIV-1/equine infectious anaemia virus TAR chimeras, we show that in addition to the well-characterized interaction with the bulge, Tat recognizes the distal stem and the loop of TAR. These data support previously proposed, but unproven, molecular models.


2005 ◽  
Vol 62 (3) ◽  
pp. 355-361 ◽  
Author(s):  
O. Chaloin ◽  
J. -C. Peter ◽  
J. -P. Briand ◽  
B. Masquida ◽  
C. Desgranges ◽  
...  
Keyword(s):  
Tar Rna ◽  

Author(s):  
Awadh Alanazi ◽  
Andrey Ivanov ◽  
Namita Kumari ◽  
Xionghao Lin ◽  
Songping Wang ◽  
...  

HIV-1 Tat protein interacts with TAR RNA and recruits CDK9/cyclin T1 and other host factors to induce HIV-1 transcription. Thus Tat-TAR RNA interaction, which is unique for HIV-1, represents an attractive target for anti-HIV-1 therapeutics. To target Tat-TAR RNA interaction, we used a crystal structure of TAR RNA with acetylpromazine bound to the bulge of TAR RNA, to dock compounds from Enamine database containing 1.6 million individual compounds. Docking identified 173 compounds that were analyzed for the inhibition of HIV-1 infection. Top ten inhibitory compounds with IC50 ≤ 6 µM were selected and the three least toxic compounds, T6780107 (IC50=2.97 μM), T0516-4834 (IC50=0.2 μM) and T5628834 (IC50=3.46 μM), were further tested for HIV-1 transcription inhibition. Only T0516-4834 compound showed selective inhibition of Tat-induced HIV-1 transcription, whereas T6780107 compound inhibited equally basal and Tat-induced transcription and T5628834 compound only inhibited basal HIV-1 transcription. The T0516-4834 compound also showed strongest inhibition of HIV-1 gag RNA expression and p24 production in CEM T cells infected with HIV-1 IIIB. Of the three compounds, only the T0516-4834 compound disrupted Tat-TAR RNA interaction indicating that it might target TAR RNA. Also, of the three tested compounds, T5628834 but not T6780107 or T0516-4834 disrupted Tat-CDK9/cyclin T1 interaction. Taken together, our study identified novel compound T0516-4834 that disrupted Tat-TAR RNA interaction and inhibited Tat-induced transcription and HIV-1 infection suggesting that this compound might serve as a new lead for anti-HIV-1 therapeutics.


1990 ◽  
Vol 10 (8) ◽  
pp. 3884-3895 ◽  
Author(s):  
R M Luche ◽  
R Sumrada ◽  
T G Cooper

Induction of the arginase (CAR1) gene expression in Saccharomyces cerevisiae has previously been shown to require participation of a cis-dominantly regulated upstream repression sequence (URS). Deletion of this element results in high-level expression of the CAR1 gene without inducer. To determine the structure of the CAR1 URS element, we performed a saturation mutagenesis. Results of the mutagenic analysis indicated that the CAR1 URS was a 9-base-pair palindromic sequence, 5'-AGCCGCCGA-3'. A DNA fragment containing this sequence was shown to bind one or more proteins by a gel shift assay. DNA fragments containing point mutations that completely eliminated URS function were not effective competitors in this assay, whereas those which supported URS function were effective competitors. Sequences in the 5'-flanking regions of 14 other genes were found to be homologous to the CAR1 URS. These sequences were shown to support varying degrees of URS function in the expression vector assay, to bind protein as demonstrated by the gel shift assay, and to compete with a DNA fragment containing the CAR1 URS for protein binding. These results indicate that the CAR1 URS element possesses the characteristics of a repressor binding site. Further, they are consistent with the suggestion that sites homologous to the CAR1 URS may be situated in the 5'-flanking regions of multiple unrelated yeast genes. The widespread occurrence of this element raises the possibility that it is the target site for one or more negatively acting general transcription factors.


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