A New Generation of Functional Tagged Proteins for HIV Fluorescence Imaging
During the last decade, there was a marked increase in the development of tools and techniques to study the molecular mechanisms of the HIV replication cycle by using fluorescence microscopy. Researchers often apply the fusion of tags and fluorophores to viral proteins, surrogate proteins, or dyes t...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-02-01
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| Series: | Viruses |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1999-4915/13/3/386 |
| _version_ | 1827604930002681856 |
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| author | João I. Mamede Joseph Griffin Stéphanie Gambut Thomas J. Hope |
| author_facet | João I. Mamede Joseph Griffin Stéphanie Gambut Thomas J. Hope |
| author_sort | João I. Mamede |
| collection | DOAJ |
| description | During the last decade, there was a marked increase in the development of tools and techniques to study the molecular mechanisms of the HIV replication cycle by using fluorescence microscopy. Researchers often apply the fusion of tags and fluorophores to viral proteins, surrogate proteins, or dyes to follow individual virus particles while they progress throughout infection. The inclusion of such fusion motifs or surrogates frequently disrupts viral infectivity or results in a change of the wild-type phenotype. Here, we detail the construction and functional characterization of two new constructs where we fused fluorescent proteins to the N-terminus of HIV-1 Integrase. In the first, IN is recruited into assembling particles via a codon optimized Gag to complement other viral constructs, while the second is fused to a Gag-Pol expression vector fully capable of integration. Our data shows that N-terminal tagged IN is functional for integration by both recovery of integration of catalytically inactive IN and by the successful infectivity of viruses carrying only labeled IN. These tools will be important to study the individual behavior of viral particles and associate such behavior to infectivity. |
| first_indexed | 2024-03-09T06:10:16Z |
| format | Article |
| id | doaj.art-3db2e5a561e944638d6c68171069bf22 |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-09T06:10:16Z |
| publishDate | 2021-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-3db2e5a561e944638d6c68171069bf222023-12-03T11:58:50ZengMDPI AGViruses1999-49152021-02-0113338610.3390/v13030386A New Generation of Functional Tagged Proteins for HIV Fluorescence ImagingJoão I. Mamede0Joseph Griffin1Stéphanie Gambut2Thomas J. Hope3Department of Microbial Pathogens and Immunity, Rush University Medical Center, Chicago, IL 60612, USADepartment of Cell and Developmental Biology, Northwestern University, Chicago, IL 60611, USADepartment of Microbial Pathogens and Immunity, Rush University Medical Center, Chicago, IL 60612, USADepartment of Cell and Developmental Biology, Northwestern University, Chicago, IL 60611, USADuring the last decade, there was a marked increase in the development of tools and techniques to study the molecular mechanisms of the HIV replication cycle by using fluorescence microscopy. Researchers often apply the fusion of tags and fluorophores to viral proteins, surrogate proteins, or dyes to follow individual virus particles while they progress throughout infection. The inclusion of such fusion motifs or surrogates frequently disrupts viral infectivity or results in a change of the wild-type phenotype. Here, we detail the construction and functional characterization of two new constructs where we fused fluorescent proteins to the N-terminus of HIV-1 Integrase. In the first, IN is recruited into assembling particles via a codon optimized Gag to complement other viral constructs, while the second is fused to a Gag-Pol expression vector fully capable of integration. Our data shows that N-terminal tagged IN is functional for integration by both recovery of integration of catalytically inactive IN and by the successful infectivity of viruses carrying only labeled IN. These tools will be important to study the individual behavior of viral particles and associate such behavior to infectivity.https://www.mdpi.com/1999-4915/13/3/386fluorescent HIVintegration competent tagged virusesHIV early-steps |
| spellingShingle | João I. Mamede Joseph Griffin Stéphanie Gambut Thomas J. Hope A New Generation of Functional Tagged Proteins for HIV Fluorescence Imaging Viruses fluorescent HIV integration competent tagged viruses HIV early-steps |
| title | A New Generation of Functional Tagged Proteins for HIV Fluorescence Imaging |
| title_full | A New Generation of Functional Tagged Proteins for HIV Fluorescence Imaging |
| title_fullStr | A New Generation of Functional Tagged Proteins for HIV Fluorescence Imaging |
| title_full_unstemmed | A New Generation of Functional Tagged Proteins for HIV Fluorescence Imaging |
| title_short | A New Generation of Functional Tagged Proteins for HIV Fluorescence Imaging |
| title_sort | new generation of functional tagged proteins for hiv fluorescence imaging |
| topic | fluorescent HIV integration competent tagged viruses HIV early-steps |
| url | https://www.mdpi.com/1999-4915/13/3/386 |
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