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Updated: Apr 11, 2026
Author Spotlight: Exploring the Role of Unfolded Protein Response in HIV-1 Replication and Infectivity
Published on: June 14, 2024
Establishment of HIV-1 model cell line GHOST(3) with stable DRiP78 and NHERF1 knockdown
Lin Zhang1, Xu-He Huang2, Ping-Ping Zhou2
1Center for Translational Medicine, Huaihe Clinical Institute, Henan University, Kaifeng 475000, China.
Abstract:
Chemokine receptors CXCR4 and CCR5 are indispensable co-receptors for HIV-1 entry into host cells. In our previous study, we identified that dopamine receptor-interacting protein 78 (DRiP78) and Na(+)-H(+) exchanger regulatory factor 1 (NHERF1) are the CXCR4 and CCR5 homo- or hetero-dimer-interacting proteins. DRiP78 and NHERF1 are able to influence the co-receptor internalization and intracellular trafficking. Over-expression of NHERF1 affects the ligands or HIV-1 gp120-induced CCR5 internalization and HIV-1 production. It is reasonable to speculate that DRiP78 and NHERF1, as well as the signaling pathways involved in viral replication, would probably affect HIV-1 replication through regulating the co-receptors. In this present study, we designed two short hairpin RNAs (shRNAs) targeting the DRiP78 and NHERF1, respectively, and constructed the pLenti6/BLOCK-iT-DEST lentiviral plasmids expressing DRiP78 or NHERF1 shRNA. The packaged lentiviruses were used to transduce the widely-applied HIV-1 model cell line GHOST(3). Then, cells with stable knockdown were established through selecting transduced cells with Blasticidin. This study, for the first time, reported the establishment of the GHOST(3) with DRiP78 and NHERF1 knockdown, which is the first stable cell line with HIV-1 co-receptor-interacting molecular defects.
Insights
Researchers created a new cell line to study HIV-1 entry. This cell line, GHOST(3) with dopamine receptor-interacting protein 78 (DRiP78) and Na(+)-H(+) exchanger regulatory factor 1 (NHERF1) knockdown, is the first with defects in HIV-1 co-receptor interactions.
Area of Science:
- Virology
- Cell Biology
- Molecular Medicine
Background:
- Chemokine receptors CXCR4 and CCR5 are crucial for HIV-1 entry.
- Dopamine receptor-interacting protein 78 (DRiP78) and Na(+)-H(+) exchanger regulatory factor 1 (NHERF1) interact with these co-receptors.
- DRiP78 and NHERF1 influence co-receptor trafficking and HIV-1 production.
Purpose of the Study:
- To investigate the role of DRiP78 and NHERF1 in HIV-1 replication.
- To establish a stable cell line for studying HIV-1 co-receptor interactions.
- To create a model for analyzing molecular defects in HIV-1 co-receptor pathways.
Main Methods:
- Designed short hairpin RNAs (shRNAs) targeting DRiP78 and NHERF1.
- Constructed lentiviral plasmids for shRNA expression.
- Transduced GHOST(3) cells and selected for stable knockdown using Blasticidin.
Main Results:
- Successfully established GHOST(3) cells with stable knockdown of DRiP78 and NHERF1.
- Created the first stable cell line with defects in HIV-1 co-receptor-interacting molecules.
- This cell line serves as a novel model for HIV-1 research.
Conclusions:
- The developed GHOST(3) cell line with DRiP78 and NHERF1 knockdown provides a unique tool for studying HIV-1 entry mechanisms.
- This model allows for the investigation of how co-receptor-interacting proteins affect viral replication.
- Further research can utilize this cell line to explore potential therapeutic targets for HIV-1.
