Part:BBa_K1965040

SbMVp

Introduction

Soybean mosaic virus protease (SbMVp) is a 27kDa protease from soybean mosaic virus (SbMV). SbMVp is also known as the nuclear inclusion a protein (NIa) and is one of the three viral proteases responsible for the processing of viral polyprotein to at least ten functional segments ^{[1, 3]}. SbMVp has been recently studied by Seo et al. as a tool for protein-protein interaction studies in the soybean. The substrate (SbMVs) has been determined to be the sequence ESVSLQ-S ^{[3, 4]}

Characterization

SbMVp has a well-defined seven amino acid recognition motif SbMVs which is defined by the amino acid sequence ESVSLQ-S ^{[3, 4]} UNIPROT: Q90069. We included that sequence into the cyclic luciferase (cycLuc_SbMVs) reporter construct.

This construct was expressed under the CMV promoter. The coding sequence for SbMVp was deposited in pSB1C3.

SbMVp was used as one of the new orthogonal proteases in our protease based signaling platform. 1 shows that SbMVp successfully cleaves its substrate and is orthogonal to other proteases (TEVp, PPVp and SuMMVp).

Figure 1. Protease orthogonality
HEK293T cells were transfected with 100ng of the indicated cycLuc reporters and 70ng of the indicated proteases. Luciferase activity was detected only in the presence of a protease and cycLuc reporter containing appropriate protease cleavage site.

References

^[1]Adams, M. J., Antoniw, J. F., & Beaudoin, F. (2005). Overview and analysis of the polyprotein cleavage sites in the family Potyviridae. Molecular Plant Pathology, 6(4), 471–487. https://doi.org/10.1111/j.1364-3703.2005.00296.x
^[2]Cho, E.-K., & Goodman, R. M. (1979). Strains of Soybean mosaic virus: classification based on virulence in resistant soybean cultivars. Phytopathology.
^[3]Seo, J.-K., Choi, H.-S., & Kim, K.-H. (2016). Engineering of soybean mosaic virus as a versatile tool for studying protein–protein interactions in soybean. Scientific Reports, 6(November 2015), 22436. https://doi.org/10.1038/srep22436
^[4]Yoon, H. Y., Hwang, D. C., Choi, K. Y., & Song, B. D. (n.d.). Proteolytic Processing of Oligopeptides Containing the Target Sequences by the Recombinant Tobacco Vein Mottling Virus NIa Proteinase. Molecules and Cells, 10(2), 213–219. Retrieved from http://cat.inist.fr/?aModele=afficheN&cpsidt=22307233

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 BsaI.rc site found at 91
  Illegal SapI.rc site found at 357