Publications
2019
Krishnan A, Pillai V, Chameettachal A, Ali L M, Pitchai F Nuzra Nagoor, Tariq S, Mustafa F, Marquet R, Rizvi T A
Purification and Functional Characterization of a Biologically Active Full-Length Feline Immunodeficiency Virus (FIV) Pr50Gag. Article de journal
Dans: Viruses, vol. 11, no. 8, p. 689, 2019, ISBN: 31357656.
Résumé | Liens | BibTeX | Étiquettes: Gag protein purification His-tag fusion protein Pr50Gag protein expression feline immunodeficiency virus (FIV) retroviral RNA packaging viral assembly, MARQUET, PAILLART, Unité ARN
@article{,
title = {Purification and Functional Characterization of a Biologically Active Full-Length Feline Immunodeficiency Virus (FIV) Pr50^{Gag}.},
author = {A Krishnan and V Pillai and A Chameettachal and L M Ali and F Nuzra Nagoor Pitchai and S Tariq and F Mustafa and R Marquet and T A Rizvi},
url = {https://www.ncbi.nlm.nih.gov/pubmed/31357656?report=&dispmax=200&tool=PubCrawler_2.23},
doi = {10.3390/v11080689},
isbn = {31357656},
year = {2019},
date = {2019-01-01},
journal = {Viruses},
volume = {11},
number = {8},
pages = {689},
abstract = {The feline immunodeficiency virus (FIV) full-length Pr50Gag precursor is a key player in the assembly of new viral particles. It is also a critical component of the efficient selection and packaging of two copies of genomic RNA (gRNA) into the newly formed virus particles from a wide pool of cellular and spliced viral RNA. To understand the molecular mechanisms involved during FIV gRNA packaging, we expressed the His6-tagged and untagged recombinant FIV Pr50Gag protein both in eukaryotic and prokaryotic cells. The recombinant Pr50Gag-His6-tag fusion protein was purified from soluble fractions of prokaryotic cultures using immobilized metal affinity chromatography (IMAC). This purified protein was able to assemble in vitro into virus-like particles (VLPs), indicating that it preserved its ability to oligomerize/multimerize. Furthermore, VLPs formed in eukaryotic cells by the FIV full-length Pr50Gag both in the presence and absence of His6-tag could package FIV sub-genomic RNA to similar levels, suggesting that the biological activity of the recombinant full-length Pr50Gag fusion protein was retained in the presence of His6-tag at the carboxy terminus. Successful expression and purification of a biologically active, recombinant full-length Pr50Gag-His6-tag fusion protein will allow study of the intricate RNA-protein interactions involved during FIV gRNA encapsidation.},
keywords = {Gag protein purification His-tag fusion protein Pr50Gag protein expression feline immunodeficiency virus (FIV) retroviral RNA packaging viral assembly, MARQUET, PAILLART, Unité ARN},
pubstate = {published},
tppubtype = {article}
}
The feline immunodeficiency virus (FIV) full-length Pr50Gag precursor is a key player in the assembly of new viral particles. It is also a critical component of the efficient selection and packaging of two copies of genomic RNA (gRNA) into the newly formed virus particles from a wide pool of cellular and spliced viral RNA. To understand the molecular mechanisms involved during FIV gRNA packaging, we expressed the His6-tagged and untagged recombinant FIV Pr50Gag protein both in eukaryotic and prokaryotic cells. The recombinant Pr50Gag-His6-tag fusion protein was purified from soluble fractions of prokaryotic cultures using immobilized metal affinity chromatography (IMAC). This purified protein was able to assemble in vitro into virus-like particles (VLPs), indicating that it preserved its ability to oligomerize/multimerize. Furthermore, VLPs formed in eukaryotic cells by the FIV full-length Pr50Gag both in the presence and absence of His6-tag could package FIV sub-genomic RNA to similar levels, suggesting that the biological activity of the recombinant full-length Pr50Gag fusion protein was retained in the presence of His6-tag at the carboxy terminus. Successful expression and purification of a biologically active, recombinant full-length Pr50Gag-His6-tag fusion protein will allow study of the intricate RNA-protein interactions involved during FIV gRNA encapsidation.