A novel aminoacid determinant of HIV-1 restriction in the TRIM5α variable 1 region isolated in a random mutagenic screen.
Human-derived antiretroviral transgenes are of great biomedical interest and are actively pursued. HIV-1 is efficiently inhibited at post-entry, pre-integration replication stages by point mutations in the variable region 1 (v1) of the human restriction factor TRIM5α. Here we use a mutated megaprimer approach to create a mutant library of TRIM5αHu v1 and to isolate a mutation at Gly330 (G330E) that inhibits transduction of an HIV-1 vector as efficiently as the previously described mutants at positions Arg332 and Arg335. As was the case for these other mutations, modification of the local v1 charge toward increased acidity was key to inhibiting HIV-1. G330E TRIM5αHu also disrupted replication-competent HIV-1 propagation in a human T cell line. Interestingly, G330E did not enhance restriction of HIV-1 when combined with mutations at Arg332 or Arg335. Accordingly, the triple mutant G330E-R332G-R335G bound purified recombinant HIV-1 capsid tubes less efficiently than the double mutant R332G-R335G did. In a structural model of the TRIM5αHu PRYSPRY domain, the addition of G330E to the double mutant R332G-R335G caused extensive changes to the capsid-binding surface, which may explain why the triple mutant was no more restrictive than the double mutant. The HIV-1 inhibitory potential of Gly330 mutants was not predicted by examination of natural TRIM5α orthologs that are known to strongly inhibit HIV-1. This work underlines the potential of random mutagenesis to isolate novel variants of human proteins with antiviral properties.
Laboratory of Retrovirology, Department of Medical Biology, Université du Québec à Trois-Rivières, 3351 Boulevard des Forges, CP500, Trois-Rivières, QC, G9A 5H7, Canada.
Quang Toan
QT
Pham
Laboratory of Retrovirology, Department of Medical Biology, Université du Québec à Trois-Rivières, 3351 Boulevard des Forges, CP500, Trois-Rivières, QC, G9A 5H7, Canada.
Maxime
M
Veillette
Department of Microbiology and Immunology, Albert Einstein College of Medicine, 1301 Morris Park-Price Center 501, Bronx, NY 10461, USA.
Alberto
A
Brandariz-Nuñez
Laboratory of Retrovirology, Department of Medical Biology, Université du Québec à Trois-Rivières, 3351 Boulevard des Forges, CP500, Trois-Rivières, QC, G9A 5H7, Canada.
Paulina
P
Pawlica
Laboratory of Retrovirology, Department of Medical Biology, Université du Québec à Trois-Rivières, 3351 Boulevard des Forges, CP500, Trois-Rivières, QC, G9A 5H7, Canada.
Caroline
C
Thibert-Lefebvre
Laboratory of Retrovirology, Department of Medical Biology, Université du Québec à Trois-Rivières, 3351 Boulevard des Forges, CP500, Trois-Rivières, QC, G9A 5H7, Canada.
Nadia
N
Chandonnet
Department of Microbiology and Immunology, Albert Einstein College of Medicine, 1301 Morris Park-Price Center 501, Bronx, NY 10461, USA.
Felipe
F
Diaz-Griffero
Laboratory of Retrovirology, Department of Medical Biology, Université du Québec à Trois-Rivières, 3351 Boulevard des Forges, CP500, Trois-Rivières, QC, G9A 5H7, Canada.
Lionel
L
Berthoux
D000090103:Antiviral Restriction Factors; D001665:Binding Sites; D002352:Carrier Proteins / Q000235:genetics* / Q000276:immunology*; D002460:Cell Line; D005820:Genetic Testing; D015497:HIV-1 / Q000276:immunology*; D006801:Humans; D008958:Models, Molecular; D016296:Mutagenesis; D050505:Mutant Proteins / Q000235:genetics / Q000276:immunology; D011485:Protein Binding; D011487:Protein Conformation; D013601:T-Lymphocytes / Q000821:virology; D014161:Transduction, Genetic; D000070584:Tripartite Motif Proteins; D044767:Ubiquitin-Protein Ligases; D014779:Virus Replication
D000090103:Antiviral Restriction Factors; D002352:Carrier Proteins; D050505:Mutant Proteins; D000070584:Tripartite Motif Proteins; C477346:TRIM5 protein, human; D044767:Ubiquitin-Protein Ligases