Abstract
The initial stages of HIV-1 infection involve the transport of the viral core into the nuclear compartment. The presence of the HIV-1 core in the nucleus triggers the translocation of CPSF6/CPSF5 from paraspeckles into nuclear speckles, forming puncta-like structures. While this phenomenon is well-documented, the efficiency of CPSF6 translocation to nuclear speckles upon HIV-1 infection varies depending on the type of cell used. In some human cell lines, only 1–2% of the cells translocate CPSF6 to nuclear speckles when exposed to a 95% infection rate. To address the issue that only 1–2% of cells translocate CPSF6 to nuclear speckles when a 95% infection rate is achieved, we screened several human cell lines and identified a human a cell line in which approximately 85% of the cells translocate CPSF6 to nuclear speckles when 95% infection rate is achieved. This cellular system has enabled the development of a robust fluorescence microscopy method to quantify the translocation of CPSF6 into nuclear speckles following HIV-1 infection. This assay holds the potential to support studies aimed at understanding the role of CPSF6 translocation to nuclear speckles in HIV-1 infection. Additionally, since the translocation of CPSF6 into nuclear speckles depends on the physical presence of the viral core in the nucleus, our method also serves as a reporter of HIV-1 nuclear import.
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Acknowledgments
This work was supported by an NIH grant AI087390 to F.D.-G. We are grateful to the NIH AIDS repository for providing plasmids and antibodies.
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Luchsinger, C., Diaz-Griffero, F. (2024). Detection of CPSF6 in Biomolecular Condensates as a Reporter of HIV-1 Nuclear Import. In: Prasad, V.R., Kalpana, G.V. (eds) HIV Protocols . Methods in Molecular Biology, vol 2807. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3862-0_9
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DOI: https://doi.org/10.1007/978-1-0716-3862-0_9
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