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Quantification of the HIV transcriptional activator complex in live cells by image‐based protein–protein interaction analysis
Author(s) -
Asamitsu Kaori,
Omagari Katsumi,
Okuda Tomoya,
Hibi Yurina,
Okamoto Takashi
Publication year - 2016
Publication title -
genes to cells
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.912
H-Index - 115
eISSN - 1365-2443
pISSN - 1356-9597
DOI - 10.1111/gtc.12375
Subject(s) - p tefb , biology , elongation factor , cyclin dependent kinase 9 , rna polymerase ii , microbiology and biotechnology , protein–protein interaction , transcription (linguistics) , transcription factor , activator (genetics) , rna , protein kinase a , kinase , cyclin dependent kinase 2 , biochemistry , promoter , ribosome , gene , gene expression , linguistics , philosophy
The virus‐encoded Tat protein is essential for HIV transcription in infected cells. The interaction of Tat with the cellular transcription elongation factor P‐TEFb (positive transcriptional elongation factor b) containing cyclin T1 (CycT1) and cyclin‐dependent kinase 9 (CDK9) is critical for its activity. In this study, we use the Fluoppi (fluorescent‐based technology detecting protein–protein interaction) system, which enables the quantification of interactions between biomolecules, such as proteins, in live cells. Quantitative measurement of the molecular interactions among Tat, CycT1 and CDK9 has showed that any third molecule enhances the binding between the other two molecules. These findings suggest that each component of the Tat:P‐TEFb complex stabilizes the overall complex, thereby supporting the efficient transcriptional elongation during viral RNA synthesis. These interactions may serve as appropriate targets for novel anti‐HIV therapy.