Limits...
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.

Show MeSH

Related in: MedlinePlus

MT4 cells were infected with recombinant SeV expressing Rh (white circles) or CM SPRY(-) (white squares) TRIM5α. Nine hours after infection, cells were superinfected with GH/SCA (A), GH/NSCG (B) or GH/SCA derivatives (C-G). Culture supernatants were separately assayed for levels of p25. Error bars show actual fluctuations between levels of p25 in duplicate samples. A representative of two independent experiments is shown.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC2944288&req=5

Figure 3: MT4 cells were infected with recombinant SeV expressing Rh (white circles) or CM SPRY(-) (white squares) TRIM5α. Nine hours after infection, cells were superinfected with GH/SCA (A), GH/NSCG (B) or GH/SCA derivatives (C-G). Culture supernatants were separately assayed for levels of p25. Error bars show actual fluctuations between levels of p25 in duplicate samples. A representative of two independent experiments is shown.

Mentions: To confirm that CA contains all determinants for restriction by Rh TRIM5α, we constructed a chimeric GH123 containing the whole region of SIVmac239 CA (GH/SCA). This virus could grow in the presence and absence of Rh TRIM5α (Figures 1 and 3A), clearly excluding the possibility that some of the determinants lie outside the CA. We then generated a chimeric GH123 containing the N-terminal half (from the 1st to 120th) of SIVmac239 CA (GH/NSCG) to further narrow down the determinant for restriction by Rh TRIM5α. Although GH/NSCG grew to lower titers than GH/SCA, even in the absence of Rh TRIM5α, this virus could also grow in the presence of Rh TRIM5α (Figures 1 and 3B). These results suggest that the N-terminal half of SIVmac239 CA is almost sufficient to evade Rh TRIM5α, even though the 179th amino acid of the C-terminal half possessed a slight effect of restriction.


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)

MT4 cells were infected with recombinant SeV expressing Rh (white circles) or CM SPRY(-) (white squares) TRIM5α. Nine hours after infection, cells were superinfected with GH/SCA (A), GH/NSCG (B) or GH/SCA derivatives (C-G). Culture supernatants were separately assayed for levels of p25. Error bars show actual fluctuations between levels of p25 in duplicate samples. A representative of two independent experiments is shown.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 3: MT4 cells were infected with recombinant SeV expressing Rh (white circles) or CM SPRY(-) (white squares) TRIM5α. Nine hours after infection, cells were superinfected with GH/SCA (A), GH/NSCG (B) or GH/SCA derivatives (C-G). Culture supernatants were separately assayed for levels of p25. Error bars show actual fluctuations between levels of p25 in duplicate samples. A representative of two independent experiments is shown.
Mentions: To confirm that CA contains all determinants for restriction by Rh TRIM5α, we constructed a chimeric GH123 containing the whole region of SIVmac239 CA (GH/SCA). This virus could grow in the presence and absence of Rh TRIM5α (Figures 1 and 3A), clearly excluding the possibility that some of the determinants lie outside the CA. We then generated a chimeric GH123 containing the N-terminal half (from the 1st to 120th) of SIVmac239 CA (GH/NSCG) to further narrow down the determinant for restriction by Rh TRIM5α. Although GH/NSCG grew to lower titers than GH/SCA, even in the absence of Rh TRIM5α, this virus could also grow in the presence of Rh TRIM5α (Figures 1 and 3B). These results suggest that the N-terminal half of SIVmac239 CA is almost sufficient to evade Rh TRIM5α, even though the 179th amino acid of the C-terminal half possessed a slight effect of restriction.

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