Purification and Functional Characterization of a Biologically Active Full-Length Feline Immunodeficiency Virus (FIV) Pr50

Overview

Journal Viruses

Publisher MDPI

Specialty Microbiology

Date 2019 Jul 31

PMID 31357656

Citations 6

Authors

Anjana Krishnan

Vineeta N Pillai

Akhil Chameettachal

Lizna Mohamed Ali

Fathima Nuzra Nagoor Pitchai

Saeed Tariq

Farah Mustafa

Roland Marquet

Tahir A Rizvi

Affiliations

Soon will be listed here.

Abstract

The feline immunodeficiency virus (FIV) full-length Pr50 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 His-tagged and untagged recombinant FIV Pr50 protein both in eukaryotic and prokaryotic cells. The recombinant Pr50-His-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 Pr50 both in the presence and absence of His-tag could package FIV sub-genomic RNA to similar levels, suggesting that the biological activity of the recombinant full-length Pr50 fusion protein was retained in the presence of His-tag at the carboxy terminus. Successful expression and purification of a biologically active, recombinant full-length Pr50-His-tag fusion protein will allow study of the intricate RNA-protein interactions involved during FIV gRNA encapsidation.

Citing Articles

MMTV RNA packaging requires an extended long-range interaction for productive Gag binding to packaging signals.

Prabhu S, Pillai V, Ali L, Vivet-Boudou V, Chameettachal A, Bernacchi S PLoS Biol. 2024; 22(10):e3002827.

PMID: 39361708 PMC: 11449360. DOI: 10.1371/journal.pbio.3002827.

Identification of a putative Gag binding site critical for feline immunodeficiency virus genomic RNA packaging.

Krishnan A, Ali L, Prabhu S, Pillai V, Chameettachal A, Vivet-Boudou V RNA. 2023; 30(1):68-88.

PMID: 37914398 PMC: 10726167. DOI: 10.1261/rna.079840.123.

Expression, purification, and functional characterization of soluble recombinant full-length simian immunodeficiency virus (SIV) Pr55.

Pillai V, Ali L, Prabhu S, Krishnan A, Tariq S, Mustafa F Heliyon. 2023; 9(1):e12892.

PMID: 36685375 PMC: 9853374. DOI: 10.1016/j.heliyon.2023.e12892.

Review and Perspectives on the Structure-Function Relationships of the Gag Subunits of Feline Immunodeficiency Virus.

Long M, Toesca J, Guillon C Pathogens. 2021; 10(11).

PMID: 34832657 PMC: 8621984. DOI: 10.3390/pathogens10111502.

A purine loop and the primer binding site are critical for the selective encapsidation of mouse mammary tumor virus genomic RNA by Pr77Gag.

Chameettachal A, Vivet-Boudou V, Pitchai F, Pillai V, Ali L, Krishnan A Nucleic Acids Res. 2021; 49(8):4668-4688.

PMID: 33836091 PMC: 8096270. DOI: 10.1093/nar/gkab223.

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