Part:BBa_K174002

Arabinose controlled protein degradation device

The purpose of this part is to selectively control the rate of degradation of proteins in Bacillus subtilis.

Griffith and Grossman, 2008 [1], showed that by tagging proteins with ssrA tags they can be tagged for recruitment by ClpX and subject to proteolysis.

Mutations on ssrA degradation tags weaken the recognition of the tagged protein by ClpX. Modified versions require SspB adaptor protein for transport to ClpX and subsequent degradation. Hence when SspB protein is expressed, the proteins tagged with modified version of ssrA tag are targeted for degradation, otherwise they remain stable [1].

This device is inducible by arabinose, therefore we can alter the arabinose levels to regulate the degradation of proteins tagged with the modified version of ssrA degradation tags.

The wild type E. coli ssrA tag is AANDENY-ALAA (SspB recognition site – ClpX recognition site).

As an example of the tag, we constructed Part:BBa_K174003 with Hin protein which was tagged with the modified version of ssrA tag below.

AANDENY-SENY-ALGG (SspB recognition site – SENY +4 Linker - ClpX recognition site)

However, it should also be noted that different degradation tags may have effect on the activity of different proteins [1]. For more information about this part, go to Newcastle iGEM 2009 [http://2009.igem.org/Team:Newcastle/Stochasticity Stochasticity] and [http://2009.igem.org/Team:Newcastle/Modelling Modelling] pages.

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]

References

1. Griffith, K. L., and A. D. Grossman. 2008. Inducible protein degradation in Bacillus subtilis using heterologous peptide tags and adaptor proteins to target substrates to the protease ClpXP. Mol. Microbiol. 70:1012-1025.