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| - | '''Unreleased structure''' | |
| | | | |
| - | The entry 6qoz is ON HOLD until Paper Publication
| + | ==CryoEM reconstruction of Cowpea Mosaic Virus (CPMV) bound to an Affimer reagent== |
| | + | <StructureSection load='6qoz' size='340' side='right'caption='[[6qoz]], [[Resolution|resolution]] 3.40Å' scene=''> |
| | + | == Structural highlights == |
| | + | <table><tr><td colspan='2'>[[6qoz]] is a 9 chain structure. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=6QOZ OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=6QOZ FirstGlance]. <br> |
| | + | </td></tr><tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=6qoz FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=6qoz OCA], [http://pdbe.org/6qoz PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=6qoz RCSB], [http://www.ebi.ac.uk/pdbsum/6qoz PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=6qoz ProSAT]</span></td></tr> |
| | + | </table> |
| | + | == Function == |
| | + | [[http://www.uniprot.org/uniprot/POL2_CPMVS POL2_CPMVS]] Movement protein: transports viral genome to neighboring plant cells directly through plasmosdesmata, without any budding. The movement protein allows efficient cell to cell propagation, by bypassing the host cell wall barrier. Acts by forming a tubular structure at the host plasmodesmata, enlarging it enough to allow free passage of virion capsids. Binds to GTP and to single-stranded RNA and single-stranded DNA in a non-sequence-specific manner.<ref>PMID:10049828</ref> <ref>PMID:15483261</ref> <ref>PMID:15165817</ref> <ref>PMID:14722313</ref> The cleavable C-terminus of small coat protein seems to be involved in the packaging of the virion RNAs. Also seems to act as suppressor of post-transcriptional gene silencing (PTGS), a mechanism of plant viral defense that limits the accumulation of viral RNAs.<ref>PMID:10049828</ref> <ref>PMID:15483261</ref> <ref>PMID:15165817</ref> <ref>PMID:14722313</ref> |
| | + | <div style="background-color:#fffaf0;"> |
| | + | == Publication Abstract from PubMed == |
| | + | Plant viruses can cause devastating losses to agriculture and are therefore a major threat to food security. The rapid identification of virally-infected crops allowing containment is essential to limit such threats, but plant viral diseases can be extremely challenging to diagnose. An ideal method for plant virus diagnosis would be a device which can be implemented easily in the field. Such devices require a binding reagent that is specific for the virus of interest. We chose to investigate the use of Affimer reagents, artificial binding proteins and a model plant virus Cowpea Mosaic virus (CPMV) empty virus like particles (eVLPs). CPMV-eVLP mimic the morphology of wild-type (WT) CPMV but lack any infectious genomic material and so do not have biocontainment issues. We have produced and purified an Affimer reagent selected for its ability to bind to CPMV-eVLP and have shown that the selected Affimer also specifically binds to WT CPMV. We have produced a 3.4 A structure of WT CPMV bound to the Affimer using cryo-electron microscopy. Finally, we have shown that this Affimer is capable of reliably detecting the virus in crude extracts of CPMV-infected leaves and can therefore form the basis for the future development of diagnostic tests. |
| | | | |
| - | Authors: Hesketh, E.L., Tiede, C., Adamson, H., Adams, T.L., Byrne, M.J., Meshcheriakova, Y., Lomonossoff, G.P., Kruse, I., McPherson, M.J., Tomlinson, D.C., Ranson, N.A.
| + | Affimer reagents as tools in diagnosing plant virus diseases.,Hesketh EL, Tiede C, Adamson H, Adams TL, Byrne MJ, Meshcheriakova Y, Kruse I, McPherson MJ, Lomonossoff GP, Tomlinson DC, Ranson NA Sci Rep. 2019 May 17;9(1):7524. doi: 10.1038/s41598-019-43945-6. PMID:31101847<ref>PMID:31101847</ref> |
| | | | |
| - | Description: CryoEM reconstructuion of Cowpea Mosaic Virus (CPMV) bound to an Affimer reagent
| + | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> |
| - | [[Category: Unreleased Structures]]
| + | </div> |
| - | [[Category: Adams, T.L]]
| + | <div class="pdbe-citations 6qoz" style="background-color:#fffaf0;"></div> |
| - | [[Category: Mcpherson, M.J]]
| + | == References == |
| - | [[Category: Hesketh, E.L]]
| + | <references/> |
| - | [[Category: Tiede, C]]
| + | __TOC__ |
| - | [[Category: Ranson, N.A]]
| + | </StructureSection> |
| - | [[Category: Kruse, I]]
| + | [[Category: Large Structures]] |
| - | [[Category: Tomlinson, D.C]]
| + | [[Category: Adams, T L]] |
| - | [[Category: Byrne, M.J]] | + | |
| - | [[Category: Lomonossoff, G.P]] | + | |
| | [[Category: Adamson, H]] | | [[Category: Adamson, H]] |
| | + | [[Category: Byrne, M J]] |
| | + | [[Category: Hesketh, E L]] |
| | + | [[Category: Kruse, I]] |
| | + | [[Category: Lomonossoff, G P]] |
| | + | [[Category: McPherson, M J]] |
| | [[Category: Meshcheriakova, Y]] | | [[Category: Meshcheriakova, Y]] |
| | + | [[Category: Ranson, N A]] |
| | + | [[Category: Tiede, C]] |
| | + | [[Category: Tomlinson, D C]] |
| | + | [[Category: Affimer reagent]] |
| | + | [[Category: Antibody-like]] |
| | + | [[Category: Comoviridae]] |
| | + | [[Category: Cpmv]] |
| | + | [[Category: Picornavirale]] |
| | + | [[Category: Virus]] |
| Structural highlights
Function
[POL2_CPMVS] Movement protein: transports viral genome to neighboring plant cells directly through plasmosdesmata, without any budding. The movement protein allows efficient cell to cell propagation, by bypassing the host cell wall barrier. Acts by forming a tubular structure at the host plasmodesmata, enlarging it enough to allow free passage of virion capsids. Binds to GTP and to single-stranded RNA and single-stranded DNA in a non-sequence-specific manner.[1] [2] [3] [4] The cleavable C-terminus of small coat protein seems to be involved in the packaging of the virion RNAs. Also seems to act as suppressor of post-transcriptional gene silencing (PTGS), a mechanism of plant viral defense that limits the accumulation of viral RNAs.[5] [6] [7] [8]
Publication Abstract from PubMed
Plant viruses can cause devastating losses to agriculture and are therefore a major threat to food security. The rapid identification of virally-infected crops allowing containment is essential to limit such threats, but plant viral diseases can be extremely challenging to diagnose. An ideal method for plant virus diagnosis would be a device which can be implemented easily in the field. Such devices require a binding reagent that is specific for the virus of interest. We chose to investigate the use of Affimer reagents, artificial binding proteins and a model plant virus Cowpea Mosaic virus (CPMV) empty virus like particles (eVLPs). CPMV-eVLP mimic the morphology of wild-type (WT) CPMV but lack any infectious genomic material and so do not have biocontainment issues. We have produced and purified an Affimer reagent selected for its ability to bind to CPMV-eVLP and have shown that the selected Affimer also specifically binds to WT CPMV. We have produced a 3.4 A structure of WT CPMV bound to the Affimer using cryo-electron microscopy. Finally, we have shown that this Affimer is capable of reliably detecting the virus in crude extracts of CPMV-infected leaves and can therefore form the basis for the future development of diagnostic tests.
Affimer reagents as tools in diagnosing plant virus diseases.,Hesketh EL, Tiede C, Adamson H, Adams TL, Byrne MJ, Meshcheriakova Y, Kruse I, McPherson MJ, Lomonossoff GP, Tomlinson DC, Ranson NA Sci Rep. 2019 May 17;9(1):7524. doi: 10.1038/s41598-019-43945-6. PMID:31101847[9]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
References
- ↑ Taylor KM, Spall VE, Butler PJ, Lomonossoff GP. The cleavable carboxyl-terminus of the small coat protein of cowpea mosaic virus is involved in RNA encapsidation. Virology. 1999 Mar 1;255(1):129-37. PMID:10049828
- ↑ Canizares MC, Taylor KM, Lomonossoff GP. Surface-exposed C-terminal amino acids of the small coat protein of Cowpea mosaic virus are required for suppression of silencing. J Gen Virol. 2004 Nov;85(Pt 11):3431-5. PMID:15483261 doi:http://dx.doi.org/85/11/3431
- ↑ Liu L, Grainger J, Canizares MC, Angell SM, Lomonossoff GP. Cowpea mosaic virus RNA-1 acts as an amplicon whose effects can be counteracted by a RNA-2-encoded suppressor of silencing. Virology. 2004 May 20;323(1):37-48. PMID:15165817 doi:http://dx.doi.org/10.1016/j.virol.2004.02.013
- ↑ Carvalho CM, Pouwels J, van Lent JW, Bisseling T, Goldbach RW, Wellink J. The movement protein of cowpea mosaic virus binds GTP and single-stranded nucleic acid in vitro. J Virol. 2004 Feb;78(3):1591-4. PMID:14722313
- ↑ Taylor KM, Spall VE, Butler PJ, Lomonossoff GP. The cleavable carboxyl-terminus of the small coat protein of cowpea mosaic virus is involved in RNA encapsidation. Virology. 1999 Mar 1;255(1):129-37. PMID:10049828
- ↑ Canizares MC, Taylor KM, Lomonossoff GP. Surface-exposed C-terminal amino acids of the small coat protein of Cowpea mosaic virus are required for suppression of silencing. J Gen Virol. 2004 Nov;85(Pt 11):3431-5. PMID:15483261 doi:http://dx.doi.org/85/11/3431
- ↑ Liu L, Grainger J, Canizares MC, Angell SM, Lomonossoff GP. Cowpea mosaic virus RNA-1 acts as an amplicon whose effects can be counteracted by a RNA-2-encoded suppressor of silencing. Virology. 2004 May 20;323(1):37-48. PMID:15165817 doi:http://dx.doi.org/10.1016/j.virol.2004.02.013
- ↑ Carvalho CM, Pouwels J, van Lent JW, Bisseling T, Goldbach RW, Wellink J. The movement protein of cowpea mosaic virus binds GTP and single-stranded nucleic acid in vitro. J Virol. 2004 Feb;78(3):1591-4. PMID:14722313
- ↑ Hesketh EL, Tiede C, Adamson H, Adams TL, Byrne MJ, Meshcheriakova Y, Kruse I, McPherson MJ, Lomonossoff GP, Tomlinson DC, Ranson NA. Affimer reagents as tools in diagnosing plant virus diseases. Sci Rep. 2019 May 17;9(1):7524. doi: 10.1038/s41598-019-43945-6. PMID:31101847 doi:http://dx.doi.org/10.1038/s41598-019-43945-6
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