Part:BBa_K1965041

SuMMVp

Introduction

Sunflower mild mosaic virus protease (SuMMVp) is a protease from sunflower mosaic virus (SuMMV). SuMMVp is also known as the nuclear inclusion a protein (NIa) and is one of the three viral proteases responsible for the processing of the viral polyprotein into at least ten functional segments^{[1, 2]}. SuMMVp has been used by Fernandez-Rodriguez et al. in a study of genetic circuits using potyviral proteases. The substrate (SuMMVs) has been defined as the sequence EEIHLQ-S/G ^[2].

Characterization

SuMMVp has a well-defined seven amino acid recognition motif SuMMVs which is defined by the amino acid sequence EEIHLQ-S/G ^[2] (Uniprot: M9NYB7). We included that sequence into cyclic luciferase (cycLuc_SuMMVs) reporter construct.

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

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

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]Fernandez-Rodriguez, J., & Voigt, C. A. (2016). Post-translational control of genetic circuits using Potyvirus proteases. Nucleic Acids Research, 44(13), 6493–502. https://doi.org/10.1093/nar/gkw537
^[3]Gulya, T. J., Shiel, P. J., Freeman, T., Jordan, R. L., Isakeit, T., & Berger, P. H. (2002). Host Range and Characterization of Sunflower mosaic virus. Phytopathology, 92(7), 694–702. https://doi.org/10.1094/PHYTO.2002.92.7.694

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
  COMPATIBLE WITH RFC[1000]