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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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Western blot analysis of CA and CypA in particles of GH123, SIVmac239 and their derivatives. Viral particles from HIV-1 NL43, HIV-2 GH123, SIVmac239, and their derivatives were purified by ultracentrifugation through a 20% sucrose cushion. A total of 120 ng of p24 of HIV-1, p25 of HIV-2 GH123 derivatives or p27 of SIVmac239 derivatives was applied for gel electropholesis. Cyp A (upper panel) and CA (lower panel) were visualized by Western blotting (WB) using an anti-CypA antibody and serum from a SIV-infected monkey, respectively.
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Figure 4: Western blot analysis of CA and CypA in particles of GH123, SIVmac239 and their derivatives. Viral particles from HIV-1 NL43, HIV-2 GH123, SIVmac239, and their derivatives were purified by ultracentrifugation through a 20% sucrose cushion. A total of 120 ng of p24 of HIV-1, p25 of HIV-2 GH123 derivatives or p27 of SIVmac239 derivatives was applied for gel electropholesis. Cyp A (upper panel) and CA (lower panel) were visualized by Western blotting (WB) using an anti-CypA antibody and serum from a SIV-infected monkey, respectively.

Mentions: It has been reported that CypA was incorporated into group M HIV-1, but not HIV-2 or SIVmac particles [57]. To confirm that the replacement of CA between GH123 and SIVmac239 did not augment CypA incorporation, we performed Western blot analysis of viral particles from GH123, SIVmac239, and their derivatives. As shown in Figure 4 (upper panel), CypA proteins were clearly detected in the particles of HIV-1 NL43 but not in those of GH123, GH/SCA, GH/SCA CypG or SIVmac239, although the amount of their CA proteins was almost comparable (Figure 4, lower panel). This result indicates that the replacement between GH123 and SIVmac239 did not augment their CypA incorporation ability.


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)

Western blot analysis of CA and CypA in particles of GH123, SIVmac239 and their derivatives. Viral particles from HIV-1 NL43, HIV-2 GH123, SIVmac239, and their derivatives were purified by ultracentrifugation through a 20% sucrose cushion. A total of 120 ng of p24 of HIV-1, p25 of HIV-2 GH123 derivatives or p27 of SIVmac239 derivatives was applied for gel electropholesis. Cyp A (upper panel) and CA (lower panel) were visualized by Western blotting (WB) using an anti-CypA antibody and serum from a SIV-infected monkey, respectively.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2944288&req=5

Figure 4: Western blot analysis of CA and CypA in particles of GH123, SIVmac239 and their derivatives. Viral particles from HIV-1 NL43, HIV-2 GH123, SIVmac239, and their derivatives were purified by ultracentrifugation through a 20% sucrose cushion. A total of 120 ng of p24 of HIV-1, p25 of HIV-2 GH123 derivatives or p27 of SIVmac239 derivatives was applied for gel electropholesis. Cyp A (upper panel) and CA (lower panel) were visualized by Western blotting (WB) using an anti-CypA antibody and serum from a SIV-infected monkey, respectively.
Mentions: It has been reported that CypA was incorporated into group M HIV-1, but not HIV-2 or SIVmac particles [57]. To confirm that the replacement of CA between GH123 and SIVmac239 did not augment CypA incorporation, we performed Western blot analysis of viral particles from GH123, SIVmac239, and their derivatives. As shown in Figure 4 (upper panel), CypA proteins were clearly detected in the particles of HIV-1 NL43 but not in those of GH123, GH/SCA, GH/SCA CypG or SIVmac239, although the amount of their CA proteins was almost comparable (Figure 4, lower panel). This result indicates that the replacement between GH123 and SIVmac239 did not augment their CypA incorporation ability.

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