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. 2019 Aug 21;10(1):3760.
doi: 10.1038/s41467-019-11668-x.

Divergent Engagements Between Adeno-Associated Viruses With Their Cellular Receptor AAVR

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Free PMC article

Divergent Engagements Between Adeno-Associated Viruses With Their Cellular Receptor AAVR

Ran Zhang et al. Nat Commun. .
Free PMC article

Abstract

Adeno-associated virus (AAV) receptor (AAVR) is an essential receptor for the entry of multiple AAV serotypes with divergent rules; however, the mechanism remains unclear. Here, we determine the structures of the AAV1-AAVR and AAV5-AAVR complexes, revealing the molecular details by which PKD1 recognizes AAV5 and PKD2 is solely engaged with AAV1. PKD2 lies on the plateau region of the AAV1 capsid. However, the AAV5-AAVR interface is strikingly different, in which PKD1 is bound at the opposite side of the spike of the AAV5 capsid than the PKD2-interacting region of AAV1. Residues in strands F/G and the CD loop of PKD1 interact directly with AAV5, whereas residues in strands B/C/E and the BC loop of PKD2 make contact with AAV1. These findings further the understanding of the distinct mechanisms by which AAVR recognizes various AAV serotypes and provide an example of a single receptor engaging multiple viral serotypes with divergent rules.

Conflict of interest statement

The authors declare no competing interests.

Figures

Fig. 1
Fig. 1
The cryo-EM structures. The central cross-sections through the cryo-EM maps and the rendered images of unbound AAV1 (a), AAV2 (c), and AAV5 (e) and AAVR-bound AAV1 (b), AAV2 (d), AAV5 (f) are shown in the left and right halves of each panel, respectively. The central cross-sections are shown with the icosahedral two-, three- and fivefold axes. The scale bars represent 100 Å. Depth cueing is used to indicate the radius by color (<90 Å: blue; 100–125 Å: from cyan to yellow; >140 Å: red). Icosahedral five- and threefold axes are represented by pentagons and triangles. Density for bound AAVRs is indicated by arrows in the central cross-sections
Fig. 2
Fig. 2
AAVR PKD1 binds with AAV5. a, b The structures of trimeric AAV5 capsomers in complex with AAVR are shown in ribbon representation from a top view (a) and as a covered surface from a perpendicular side view (b). The rough inner and outer boundaries of the viral capsid are marked with gray arcs in b. The three AAV5 capsomers are colored blue, green, and cyan, respectively. Bound AAVRs are shown in orange. The five- and threefold axes are indicated by pentagons and triangles, respectively. The interacting residues on one AAVR PKD1 and the corresponding AAV5 capsomers are colored red and gold, respectively, in b, and the interacting region is framed with dashed lines. The residues in PKD1 at the interface are shown in red (c); the interface residues in AAV5 capsomer A (d), and capsomer B (e) are colored gold and labeled. See also Supplementary Table 3. f A total of nine AAVR mutants were tested for their ability to bind AAV5 by BIAcore sensorgrams in triplicate experiments. The calculated KD values for the binding of each mutant to AAV5 are summarized as the mean values of three experiments with standard errors. The original curves are presented in Supplementary Fig. 6. g The impact of AAVR PKD1 mutants overexpressed in AAVR-silenced HEK293T cells on AAV5 transduction. Cells were transfected with wt AAVR or different AAVR mutants as indicated followed by infection with AAV5-mCherry at an MOI of 3 × 106 vg cell−1. The percentage of mCherry-positive cells are plotted as means ± standard errors (n = 3). Expression of wt AAVR and mutant AAVRs was evaluated by immunoblot analysis with β-actin as a control (see Supplementary Fig. 13). h AAV5 molecules bearing capsid mutations at the receptor binding sites impacted viral transduction. HEK293T cells were infected with serial dilutions of AAV5 mutants starting at an MOI of 3 × 106 vg cell−1. Functional titer refers to the number of virus particles that can infect the cell in every microliter. GFP expression was determined 48 h post-transduction. The functional titers are plotted as means ±  standard errors (n = 3). Source data are provided as a Source Data file
Fig. 3
Fig. 3
AAVR PKD2 binds with AAV1. a, b The structures of trimeric AAV1 capsomers in complex with AAVR are shown in ribbon representation from a top view (a) and as a covered surface from a perpendicular side view (b). The rough inner and outer boundaries of the shell are marked with gray arcs. The three AAV1 capsomers are colored blue, green, and cyan, respectively. Bound AAVRs are shown in magenta. The five- and threefold axes are indicated by pentagons and triangles, respectively. The interacting residues on one AAVR and the interacting AAV1 capsomers are colored blue/white and pink in b, and the interacting region is framed. The interface amino acid residues in AAVR PKD2 are shown in bluewhite in c; the interface residues in AAV2 capsomer A (d) and capsomer B (e) are labeled and colored pink. See also Supplementary Table 5. f A total of 13 AAVR mutants were tested for their ability to bind AAV5 capsid by BIAcore sensorgrams in triplicate experiments. The calculated KD values for the binding of each mutant to AAV1 are summarized. The original curves are shown in Supplementary Fig. 6. g The overexpression of AAVR PKD2 mutants in AAVR-silenced HEK293T cells impacted on AAV1 transduction. Cells were transfected with wt AAVR or different AAVR mutants as indicated followed by infection with AAV1-mCherry at an MOI of 5 × 105 vg cell−1. The percentage of mCherry-positive cells are plotted as means ±  standard errors (n = 3). Expression of wt AAVR and mutant AAVRs was evaluated by immunoblot analysis with β-actin as a control (see Supplementary Fig. 13). h AAV1 viral particles bearing capsid mutations at the receptor-binding sites impacted viral transduction. HEK293T cells were infected with serial dilutions of mutant AAV1 starting at an MOI of 5 × 105 vg cell−1. Functional titer refers to the number of virus particles that can infect the cell in every microliter. GFP expression was determined 48 h post-transduction. The functional titers are plotted as means ±  standard errors (n = 3). Source data are provided as a Source Data file
Fig. 4
Fig. 4
Comparison of the binding between receptors and AAV capsids. af Roadmap depictions of the AAV1, AAV2, and AAV5 icosahedral surfaces projected onto a plane and showing an area larger than one icosahedral face (outlined as a triangle with the three- and fivefold vertices marked with triangles and pentagons, respectively). Two angles (θ, ϕ) define a vector and thus a location on the icosahedral surface. The roadmaps are radially depth cued, as shown by the key, from blue (radius = 80 Å) to red (radius = 150 Å). The footprints of the HSPG analog and SIA, or AAVR on different AAV capsids are outlined by white, purple, red, and orange lines. gi Three structural elements of the AAV2 capsid surrounding the bound HSPG analog are shown as cartoon diagrams and colored red, whereas their counterparts in AAV1 and AAV5 are colored blue and green, respectively. The bound HSPG analog is displayed as semi-transparent sticks. j The structural elements of the AAV1 capsid surrounding the bound SIA are shown as cartoon diagrams and colored blue, whereas their counterparts in AAV2 and AAV5 are colored red and green, respectively. The bound SIA is displayed as semi-transparent sticks. k The structural elements of the AAV5 capsid forming the site A to bind SIA are shown as cartoon diagrams and colored green, whereas their counterparts in AAV1 and AAV2 are colored blue and red, respectively. The bound SIA is displayed as semi-transparent sticks. lo Four structural elements of the AAV5 capsid interacting with PKD1 and their counterparts in the AAV1 and AAV2 capsids are shown as cartoon diagrams with the same color scheme above. The bound PKD1 is represented as a semi-transparent cartoon. ps Four structural elements of the AAV1 capsid interacting with PKD2 and their counterparts in AAV2 and AAV5 are shown as cartoon diagrams with the same color scheme above. The bound PKD2 is represented as a semi-transparent cartoon

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