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Imaging of intracellular trafficking of the HIV-1 nucleocapsid protein

Abstract : The nucleocapsid protein of HIV-1 (NCp7) plays an important role in several stages of the viral cycle. Its interactions in the intracellular environment under conditions close to true infection are not well described. The objective of this thesis project was to develop imaging approaches to monitor NCp7 during the early stages of the viral cycle directly in infected cells. By analyzing the intensity of the fluorescent pseudoviruses, we showed a fluorescence "dequenching" effect indicating a decrease in the NCp7 concentration within the virus particles, most likely resulting from the release of the nucleocapsid molecules from the viral complexes. These results show for the first time the release of NCp7 into the cytoplasm during reverse transcription. The pseudoviruses NCp7-TC were then imaged in high resolution by PALM microscopy. The distribution images of labeled NCp7 revealed the presence of NCp7-TC within the nucleus. We have demonstrated the presence of NCp7-TC in the vicinity of nuclear pores by electron microscopy.
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Iryna Lysova. Imaging of intracellular trafficking of the HIV-1 nucleocapsid protein. Virology. Université de Strasbourg, 2017. English. ⟨NNT : 2017STRAJ092⟩. ⟨tel-02917956⟩

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