Difference between revisions of "Part:BBa K404010"
 
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The AAV capsid consists of 60 capsid protein subunits. The three cap proteins VP1, VP2, and VP3 are encoded in an overlapping reading frame. Arranged in a stoichiometric ratio of 1:1:10, they form an icosahedral symmetry. The mRNA encoding for the cap proteins is transcribed from p40 and alternative spliced to minor and major products. Alternative splicing and translation initiation of VP2 at a nonconventional ACG initiation codon promote the expression of VP1, VP2 and VP3. The VP proteins share a common C terminus and stop codon, but begin with a different start codon. The translation of VP2 from the major spliced mRNA is not as efficient as of VP1 and VP3 because it initiates at a Thr codon (ACG). The N terminus of VP1 has an extension of 65 amino acids and similar to VP1, it contains two functional elements: a phospholipase A2 (PLA2) domain and nuclear localization signals (BR)(+). These elements are conserved almost in all parvoviruses. The exact role of VP2 remains unknown, although the protein is thought to be nonessential for viral assembly and infectivity.(Quelle)
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The AAV capsid consists of 60 capsid protein subunits. The three cap proteins VP1, VP2, and VP3 are encoded in an overlapping reading frame. Arranged in a stoichiometric ratio of 1:1:10, they form an icosahedral symmetry. The mRNA encoding for the cap proteins is transcribed from p40 and alternative spliced to minor and major products. Alternative splicing and translation initiation of VP2 at a nonconventional ACG initiation codon promote the expression of VP1, VP2 and VP3. The VP proteins share a common C terminus and stop codon, but begin with a different start codon. The translation of VP2 from the major spliced mRNA is not as efficient as of VP1 and VP3 because it initiates at a Thr codon (ACG). The N terminus of VP2 has an extension of 65 amino acids and similar to VP1, it contains two functional elements: a phospholipase A2 (PLA2) domain and nuclear localization signals (BR)(+). These elements are conserved almost in all parvoviruses. The PLA2 domain is involved in successful infection because it needs to be presented on the surface for endosomal release. VP2 is expendable and fusion of larger motifs to its N-terminus does not affect viral assembly and genome packaging.  
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<h3>Characterization</h3>
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<h3>References</h3>
 
<h3>References</h3>
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<b>Cassinotti, P., Weitzand, M., & Tratschin, J. D.</b>, 1988. Organization of the adeno-associated virus (AAV) capsid gene: mapping of a minor spliced mRNA coding for virus capsid protein. Virology, 167(1), 176-84 <br />
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<b>DiPrimio, Asokan, Govindasamy, Agbandje-McKenna, & Samulski</b>, June 2008. Surface loop dynamics in adeno-associated virus capsid assembly. Journal of virology, 167(1), 5178–5189 <br />
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<b>Warrington, K. H., Gorbatyuk, O. S., Harrison, J. K., Opie, S. R., Zolotukhin, S., Muzyczka, N., et al. </b>(2004). Adeno-associated virus type 2 VP2 capsid protein is nonessential and can tolerate large peptide insertions at its N terminus. Journal of virology, 78(12), 6595-609. doi: 10.1128/JVI.78.12.6595-6609.2004.<br />
  
 
<center><img src="https://static.igem.org/mediawiki/parts/a/a7/Freiburg10_Cap_proteins_VP1_2%263.png" width="600"  
 
<center><img src="https://static.igem.org/mediawiki/parts/a/a7/Freiburg10_Cap_proteins_VP1_2%263.png" width="600"  

Latest revision as of 21:32, 27 October 2010

[AAV2]-VP2

AAV2-VP2
BioBrick Nr. BBa_K404010
RFC standard RFC 25
Requirement pSB1C3_001
Source pAAV_RC from Stratagene
Submitted by [http://2010.igem.org/Team:Freiburg_Bioware FreiGEM 2010]

The AAV capsid consists of 60 capsid protein subunits. The three cap proteins VP1, VP2, and VP3 are encoded in an overlapping reading frame. Arranged in a stoichiometric ratio of 1:1:10, they form an icosahedral symmetry. The mRNA encoding for the cap proteins is transcribed from p40 and alternative spliced to minor and major products. Alternative splicing and translation initiation of VP2 at a nonconventional ACG initiation codon promote the expression of VP1, VP2 and VP3. The VP proteins share a common C terminus and stop codon, but begin with a different start codon. The translation of VP2 from the major spliced mRNA is not as efficient as of VP1 and VP3 because it initiates at a Thr codon (ACG). The N terminus of VP2 has an extension of 65 amino acids and similar to VP1, it contains two functional elements: a phospholipase A2 (PLA2) domain and nuclear localization signals (BR)(+). These elements are conserved almost in all parvoviruses. The PLA2 domain is involved in successful infection because it needs to be presented on the surface for endosomal release. VP2 is expendable and fusion of larger motifs to its N-terminus does not affect viral assembly and genome packaging.

References

DiPrimio, Asokan, Govindasamy, Agbandje-McKenna, & Samulski, June 2008. Surface loop dynamics in adeno-associated virus capsid assembly. Journal of virology, 167(1), 5178–5189
Warrington, K. H., Gorbatyuk, O. S., Harrison, J. K., Opie, S. R., Zolotukhin, S., Muzyczka, N., et al. (2004). Adeno-associated virus type 2 VP2 capsid protein is nonessential and can tolerate large peptide insertions at its N terminus. Journal of virology, 78(12), 6595-609. doi: 10.1128/JVI.78.12.6595-6609.2004.
Figure 1: The VP proteins are encoded in an overlapping open reading frame. .

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 755