Abstract
Protein-protein interactions (PPIs) occur in a vast variety of cellular processes, and many processes are regulated by multiple protein interactions. Identification of PPIs is essential for the analysis of biological pathways and to further understand underlying molecular mechanisms. However, visualization and identification of multiprotein complexes, including ternary complexes in living cells under physiological conditions, remains challenging. In this work, we reported a three-fragment fluorescence complementation (TFFC) by splitting the Venus fluorescent protein for visualizing ternary complexes in living cells under physiological conditions. With this Venus-based TFFC system, we identified the multi-interaction of weak-affinity ternary complexes under physiological conditions. The TFFC system was further applied to the analysis of multi-interactions during the HIV-1 integration process, revealing the important role of the barrier-to-autointegration factor protein in HIV-1 integration. This TFFC system provides a useful tool for visualizing and identifying ternary complexes in living cells under physiological conditions.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adaptor Proteins, Signal Transducing / genetics
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Adaptor Proteins, Signal Transducing / metabolism
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Animals
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Bacterial Proteins / chemistry*
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Bacterial Proteins / genetics
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Chlorocebus aethiops
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DNA-Binding Proteins / genetics
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DNA-Binding Proteins / metabolism
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Fluorescence
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HEK293 Cells
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HIV Integrase / genetics
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HIV Integrase / metabolism
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HeLa Cells
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Humans
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Luminescent Proteins / chemistry*
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Luminescent Proteins / genetics
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Microscopy, Confocal / methods
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Microscopy, Fluorescence / methods*
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Multiprotein Complexes / analysis*
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Multiprotein Complexes / genetics
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Multiprotein Complexes / metabolism
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NFATC Transcription Factors / genetics
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NFATC Transcription Factors / metabolism
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Nuclear Proteins / genetics
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Nuclear Proteins / metabolism
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Peptide Fragments / chemistry*
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Peptide Fragments / genetics
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Protein Binding
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Protein Multimerization
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Proto-Oncogene Proteins c-fos / genetics
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Proto-Oncogene Proteins c-fos / metabolism
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Proto-Oncogene Proteins c-jun / genetics
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Proto-Oncogene Proteins c-jun / metabolism
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Transcription Factors / genetics
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Transcription Factors / metabolism
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Vero Cells
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Virus Integration / physiology
Substances
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Adaptor Proteins, Signal Transducing
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BANF1 protein, human
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Bacterial Proteins
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DNA-Binding Proteins
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FOS protein, human
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Luminescent Proteins
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Multiprotein Complexes
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NFATC Transcription Factors
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NFATC2 protein, human
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Nuclear Proteins
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PSIP1 protein, human
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Peptide Fragments
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Proto-Oncogene Proteins c-fos
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Proto-Oncogene Proteins c-jun
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Transcription Factors
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yellow fluorescent protein, Bacteria
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HIV Integrase
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p31 integrase protein, Human immunodeficiency virus 1