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Molecular determinants for recognition of divergent SAMHD1 proteins by the lentiviral accessory protein Vpx.

Schwefel D, Boucherit VC, Christodoulou E, Walker PA, Stoye JP, Bishop KN, Taylor IA - Cell Host Microbe (2015)

Bottom Line: Comparison with Vpx from SIV that infects sooty mangabeys (SIVsmm) complexed with SAMHD1-DCAF1 identifies molecular determinants directing Vpx lineages to N- or C-terminal SAMHD1 sequences.Inspection of the Vpx-DCAF1 interface also reveals conservation of Vpx with the evolutionally related HIV-1/SIV accessory protein Vpr.These data suggest a unified model for how Vpx and Vpr exploit DCAF1 to promote viral replication.

View Article: PubMed Central - PubMed

Affiliation: Division of Molecular Structure, MRC National Institute for Medical Research, The Ridgeway, Mill Hill, London NW7 1AA, UK.

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Structure of the SIVmnd-2 Vpx/DCAF1-CtD/SAMHD1mnd-NtD Complex(A) Schematic of components of the protein complex.(B) Analytical gel filtration of the in vitro-assembled ternary protein complex consisting of SIVmnd-2 Vpx, DCAF1-CtD and SAMHD1mnd-NtD (Peak-1). The inset shows SDS-PAGE analysis of the indicated peak fractions.(C) Structure of the ternary complex. The backbone for each protein is shown in cartoon representation, SIVmnd-2 Vpx (blue), DCAF1-CtD (gray), and SAMHD1mnd-NtD (magenta). See also Figure S2. The solvent-accessible surface is also shown for DCAF1-CtD, and a zinc ion co-ordinated by Vpx is displayed as a gray sphere, see also Figure S3. For SIVmnd-2 Vpx and SAMHD1mnd-NtD, residue numbers at chain termini are indicated and secondary structure elements labeled.
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fig2: Structure of the SIVmnd-2 Vpx/DCAF1-CtD/SAMHD1mnd-NtD Complex(A) Schematic of components of the protein complex.(B) Analytical gel filtration of the in vitro-assembled ternary protein complex consisting of SIVmnd-2 Vpx, DCAF1-CtD and SAMHD1mnd-NtD (Peak-1). The inset shows SDS-PAGE analysis of the indicated peak fractions.(C) Structure of the ternary complex. The backbone for each protein is shown in cartoon representation, SIVmnd-2 Vpx (blue), DCAF1-CtD (gray), and SAMHD1mnd-NtD (magenta). See also Figure S2. The solvent-accessible surface is also shown for DCAF1-CtD, and a zinc ion co-ordinated by Vpx is displayed as a gray sphere, see also Figure S3. For SIVmnd-2 Vpx and SAMHD1mnd-NtD, residue numbers at chain termini are indicated and secondary structure elements labeled.

Mentions: To gain further insights into molecular recognition in the SIVmnd-2 Vpx system, SIVmnd-2 Vpx, SAMHD1mnd-NtD, and DCAF1-CtD (Figure 2A) were assembled in vitro using recombinant proteins and the ternary complex purified by size-exclusion chromatography (Figure 2B). The protein complex crystallized in space group P6522, the crystals diffracted to 2.8 Å resolution, and the structure was solved by molecular replacement using the SIVsmm Vpx/SAMHD1hs-CtD/DCAF1-CtD structure (Schwefel et al., 2014) as a search model. Details of the data collection, structure determination, map quality, and refinement are presented in Table 1 and Figure S2.


Molecular determinants for recognition of divergent SAMHD1 proteins by the lentiviral accessory protein Vpx.

Schwefel D, Boucherit VC, Christodoulou E, Walker PA, Stoye JP, Bishop KN, Taylor IA - Cell Host Microbe (2015)

Structure of the SIVmnd-2 Vpx/DCAF1-CtD/SAMHD1mnd-NtD Complex(A) Schematic of components of the protein complex.(B) Analytical gel filtration of the in vitro-assembled ternary protein complex consisting of SIVmnd-2 Vpx, DCAF1-CtD and SAMHD1mnd-NtD (Peak-1). The inset shows SDS-PAGE analysis of the indicated peak fractions.(C) Structure of the ternary complex. The backbone for each protein is shown in cartoon representation, SIVmnd-2 Vpx (blue), DCAF1-CtD (gray), and SAMHD1mnd-NtD (magenta). See also Figure S2. The solvent-accessible surface is also shown for DCAF1-CtD, and a zinc ion co-ordinated by Vpx is displayed as a gray sphere, see also Figure S3. For SIVmnd-2 Vpx and SAMHD1mnd-NtD, residue numbers at chain termini are indicated and secondary structure elements labeled.
© Copyright Policy - CC BY
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4400269&req=5

fig2: Structure of the SIVmnd-2 Vpx/DCAF1-CtD/SAMHD1mnd-NtD Complex(A) Schematic of components of the protein complex.(B) Analytical gel filtration of the in vitro-assembled ternary protein complex consisting of SIVmnd-2 Vpx, DCAF1-CtD and SAMHD1mnd-NtD (Peak-1). The inset shows SDS-PAGE analysis of the indicated peak fractions.(C) Structure of the ternary complex. The backbone for each protein is shown in cartoon representation, SIVmnd-2 Vpx (blue), DCAF1-CtD (gray), and SAMHD1mnd-NtD (magenta). See also Figure S2. The solvent-accessible surface is also shown for DCAF1-CtD, and a zinc ion co-ordinated by Vpx is displayed as a gray sphere, see also Figure S3. For SIVmnd-2 Vpx and SAMHD1mnd-NtD, residue numbers at chain termini are indicated and secondary structure elements labeled.
Mentions: To gain further insights into molecular recognition in the SIVmnd-2 Vpx system, SIVmnd-2 Vpx, SAMHD1mnd-NtD, and DCAF1-CtD (Figure 2A) were assembled in vitro using recombinant proteins and the ternary complex purified by size-exclusion chromatography (Figure 2B). The protein complex crystallized in space group P6522, the crystals diffracted to 2.8 Å resolution, and the structure was solved by molecular replacement using the SIVsmm Vpx/SAMHD1hs-CtD/DCAF1-CtD structure (Schwefel et al., 2014) as a search model. Details of the data collection, structure determination, map quality, and refinement are presented in Table 1 and Figure S2.

Bottom Line: Comparison with Vpx from SIV that infects sooty mangabeys (SIVsmm) complexed with SAMHD1-DCAF1 identifies molecular determinants directing Vpx lineages to N- or C-terminal SAMHD1 sequences.Inspection of the Vpx-DCAF1 interface also reveals conservation of Vpx with the evolutionally related HIV-1/SIV accessory protein Vpr.These data suggest a unified model for how Vpx and Vpr exploit DCAF1 to promote viral replication.

View Article: PubMed Central - PubMed

Affiliation: Division of Molecular Structure, MRC National Institute for Medical Research, The Ridgeway, Mill Hill, London NW7 1AA, UK.

Show MeSH
Related in: MedlinePlus