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Multiple sites in the N-terminal half of simian immunodeficiency virus capsid protein contribute to evasion from rhesus monkey TRIM5α-mediated restriction.

Kono K, Song H, Yokoyama M, Sato H, Shioda T, Nakayama EE - Retrovirology (2010)

Bottom Line: In addition, the N-terminal portion (from the 5th to 12th amino acid residues) and the 107th and 109th amino acid residues in α-helix 6 of SIVmac CA are necessary for complete evasion from Rh TRIM5α-mediated restriction.A three-dimensional model of hexameric GH123 CA showed that these multiple regions are located on the CA surface, suggesting their direct interaction with TRIM5α.We found that multiple regions of the SIVmac CA are necessary for complete evasion from Rh TRIM5α restriction.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Viral Infections, Research Institute for Microbial Diseases, Osaka University, 3-1 Yamada-oka, Suita, Osaka 565-0871, Japan.

ABSTRACT

Background: We previously reported that cynomolgus monkey (CM) TRIM5α could restrict human immunodeficiency virus type 2 (HIV-2) strains carrying a proline at the 120th position of the capsid protein (CA), but it failed to restrict those with a glutamine or an alanine. In contrast, rhesus monkey (Rh) TRIM5α could restrict all HIV-2 strains tested but not simian immunodeficiency virus isolated from macaque (SIVmac), despite its genetic similarity to HIV-2.

Results: We attempted to identify the viral determinant of SIVmac evasion from Rh TRIM5α-mediated restriction using chimeric viruses formed between SIVmac239 and HIV-2 GH123 strains. Consistent with a previous study, chimeric viruses carrying the loop between α-helices 4 and 5 (L4/5) (from the 82nd to 99th amino acid residues) of HIV-2 CA were efficiently restricted by Rh TRIM5α. However, the corresponding loop of SIVmac239 CA alone (from the 81st to 97th amino acid residues) was not sufficient to evade Rh TRIM5α restriction in the HIV-2 background. A single glutamine-to-proline substitution at the 118th amino acid of SIVmac239 CA, corresponding to the 120th amino acid of HIV-2 GH123, also increased susceptibility to Rh TRIM5α, indicating that glutamine at the 118th of SIVmac239 CA is necessary to evade Rh TRIM5α. In addition, the N-terminal portion (from the 5th to 12th amino acid residues) and the 107th and 109th amino acid residues in α-helix 6 of SIVmac CA are necessary for complete evasion from Rh TRIM5α-mediated restriction. A three-dimensional model of hexameric GH123 CA showed that these multiple regions are located on the CA surface, suggesting their direct interaction with TRIM5α.

Conclusion: We found that multiple regions of the SIVmac CA are necessary for complete evasion from Rh TRIM5α restriction.

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Activity of GH123, SIVmac239, and their derivatives to saturate TRIM5α in Rh cells. (A) Rh FRhK-4 cells were pretreated with equal amounts of VSV-G pseudotyped particles (800 ng of p25 or p27) of GH123, GH123/Q, GH123/CypS, GH123/CypS 120Q, GH/NSCG, GH/SCA N-G, GH/SCA CypG, GH/SCA, SIVmac239 or SIVmac239/P for 2 h. Cells were then infected with the VSV-G pseudotyped GFP-expressing HIV-1 vector carrying SIVmac L4/5. Data from triplicate samples (means ± SD) expressed as % GFP positive cells subtracted with the value of mock-treated cells (24.88%) are shown. Statistical significance of differences was calculated using the t-test. Asterisks above bars show differences between indicated viruses and SIVmac239. ***, P < 0.001; **, P < 0.01; ns, not significant. The statistical significance of differences between GH123 and GH123/CypS and that between GH123 and GH/SCA CypG were both < 0.001.
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Figure 5: Activity of GH123, SIVmac239, and their derivatives to saturate TRIM5α in Rh cells. (A) Rh FRhK-4 cells were pretreated with equal amounts of VSV-G pseudotyped particles (800 ng of p25 or p27) of GH123, GH123/Q, GH123/CypS, GH123/CypS 120Q, GH/NSCG, GH/SCA N-G, GH/SCA CypG, GH/SCA, SIVmac239 or SIVmac239/P for 2 h. Cells were then infected with the VSV-G pseudotyped GFP-expressing HIV-1 vector carrying SIVmac L4/5. Data from triplicate samples (means ± SD) expressed as % GFP positive cells subtracted with the value of mock-treated cells (24.88%) are shown. Statistical significance of differences was calculated using the t-test. Asterisks above bars show differences between indicated viruses and SIVmac239. ***, P < 0.001; **, P < 0.01; ns, not significant. The statistical significance of differences between GH123 and GH123/CypS and that between GH123 and GH/SCA CypG were both < 0.001.

Mentions: Cells treated with HIV-2 GH123 particles showed enhanced susceptibility to HIV-1 infection compared with non-treated cells (Figure 5), demonstrating that TRIM5α in FRhK4 cells was saturated by the high dose of the particles. In contrast, cells treated with SIVmac239 particles showed very low levels of enhancement. Cells treated with particles carrying GH123/Q showed similar levels of enhanced susceptibility to HIV-1 infection to those of HIV-2 GH123, while cells treated with particles of GH123/CypS, GH123/CypS 120Q, GH/SCA CypG or SIVmac239/P showed intermediate levels of enhancement (Figure 5).


Multiple sites in the N-terminal half of simian immunodeficiency virus capsid protein contribute to evasion from rhesus monkey TRIM5α-mediated restriction.

Kono K, Song H, Yokoyama M, Sato H, Shioda T, Nakayama EE - Retrovirology (2010)

Activity of GH123, SIVmac239, and their derivatives to saturate TRIM5α in Rh cells. (A) Rh FRhK-4 cells were pretreated with equal amounts of VSV-G pseudotyped particles (800 ng of p25 or p27) of GH123, GH123/Q, GH123/CypS, GH123/CypS 120Q, GH/NSCG, GH/SCA N-G, GH/SCA CypG, GH/SCA, SIVmac239 or SIVmac239/P for 2 h. Cells were then infected with the VSV-G pseudotyped GFP-expressing HIV-1 vector carrying SIVmac L4/5. Data from triplicate samples (means ± SD) expressed as % GFP positive cells subtracted with the value of mock-treated cells (24.88%) are shown. Statistical significance of differences was calculated using the t-test. Asterisks above bars show differences between indicated viruses and SIVmac239. ***, P < 0.001; **, P < 0.01; ns, not significant. The statistical significance of differences between GH123 and GH123/CypS and that between GH123 and GH/SCA CypG were both < 0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
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Figure 5: Activity of GH123, SIVmac239, and their derivatives to saturate TRIM5α in Rh cells. (A) Rh FRhK-4 cells were pretreated with equal amounts of VSV-G pseudotyped particles (800 ng of p25 or p27) of GH123, GH123/Q, GH123/CypS, GH123/CypS 120Q, GH/NSCG, GH/SCA N-G, GH/SCA CypG, GH/SCA, SIVmac239 or SIVmac239/P for 2 h. Cells were then infected with the VSV-G pseudotyped GFP-expressing HIV-1 vector carrying SIVmac L4/5. Data from triplicate samples (means ± SD) expressed as % GFP positive cells subtracted with the value of mock-treated cells (24.88%) are shown. Statistical significance of differences was calculated using the t-test. Asterisks above bars show differences between indicated viruses and SIVmac239. ***, P < 0.001; **, P < 0.01; ns, not significant. The statistical significance of differences between GH123 and GH123/CypS and that between GH123 and GH/SCA CypG were both < 0.001.
Mentions: Cells treated with HIV-2 GH123 particles showed enhanced susceptibility to HIV-1 infection compared with non-treated cells (Figure 5), demonstrating that TRIM5α in FRhK4 cells was saturated by the high dose of the particles. In contrast, cells treated with SIVmac239 particles showed very low levels of enhancement. Cells treated with particles carrying GH123/Q showed similar levels of enhanced susceptibility to HIV-1 infection to those of HIV-2 GH123, while cells treated with particles of GH123/CypS, GH123/CypS 120Q, GH/SCA CypG or SIVmac239/P showed intermediate levels of enhancement (Figure 5).

Bottom Line: In addition, the N-terminal portion (from the 5th to 12th amino acid residues) and the 107th and 109th amino acid residues in α-helix 6 of SIVmac CA are necessary for complete evasion from Rh TRIM5α-mediated restriction.A three-dimensional model of hexameric GH123 CA showed that these multiple regions are located on the CA surface, suggesting their direct interaction with TRIM5α.We found that multiple regions of the SIVmac CA are necessary for complete evasion from Rh TRIM5α restriction.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Viral Infections, Research Institute for Microbial Diseases, Osaka University, 3-1 Yamada-oka, Suita, Osaka 565-0871, Japan.

ABSTRACT

Background: We previously reported that cynomolgus monkey (CM) TRIM5α could restrict human immunodeficiency virus type 2 (HIV-2) strains carrying a proline at the 120th position of the capsid protein (CA), but it failed to restrict those with a glutamine or an alanine. In contrast, rhesus monkey (Rh) TRIM5α could restrict all HIV-2 strains tested but not simian immunodeficiency virus isolated from macaque (SIVmac), despite its genetic similarity to HIV-2.

Results: We attempted to identify the viral determinant of SIVmac evasion from Rh TRIM5α-mediated restriction using chimeric viruses formed between SIVmac239 and HIV-2 GH123 strains. Consistent with a previous study, chimeric viruses carrying the loop between α-helices 4 and 5 (L4/5) (from the 82nd to 99th amino acid residues) of HIV-2 CA were efficiently restricted by Rh TRIM5α. However, the corresponding loop of SIVmac239 CA alone (from the 81st to 97th amino acid residues) was not sufficient to evade Rh TRIM5α restriction in the HIV-2 background. A single glutamine-to-proline substitution at the 118th amino acid of SIVmac239 CA, corresponding to the 120th amino acid of HIV-2 GH123, also increased susceptibility to Rh TRIM5α, indicating that glutamine at the 118th of SIVmac239 CA is necessary to evade Rh TRIM5α. In addition, the N-terminal portion (from the 5th to 12th amino acid residues) and the 107th and 109th amino acid residues in α-helix 6 of SIVmac CA are necessary for complete evasion from Rh TRIM5α-mediated restriction. A three-dimensional model of hexameric GH123 CA showed that these multiple regions are located on the CA surface, suggesting their direct interaction with TRIM5α.

Conclusion: We found that multiple regions of the SIVmac CA are necessary for complete evasion from Rh TRIM5α restriction.

Show MeSH
Related in: MedlinePlus