Part:BBa_K3198007
HicA-LuxABCDE
This part contains the toxin component of a type II toxin-antitoxin (TA) system. HicA is a probable translation-independent mRNA interferase.
Bacterial lux operons encodes five enzymes involved in the light-generating pathway. LuxA and LuxB encode the two subunits of the bacterial luciferase, while the products of LuxC, LuxD and LuxE synthesise the substrate for the light emitting reaction, tetradecanal. The exact function of LuxG is unknown, and it appears to be non-essential for light emission, but its presence increases light output.
Description
This part contains the toxin component of a type II toxin-antitoxin (TA) system. HicA is a probable translation-independent mRNA interferase.
Bacterial lux operons encodes five enzymes involved in the light-generating pathway. LuxA and LuxB encode the two subunits of the bacterial luciferase, while the products of LuxC, LuxD and LuxE synthesise the substrate for the light emitting reaction, tetradecanal. The exact function of LuxG is unknown, and it appears to be non-essential for light emission, but its presence increases light output.
Usage
Team NUS Singapore 2019 has added a new biobrick (BBa_K3198007) into the iGEM repository this year. This biobrick was found to possess bacteriostatic effect as reported by Gerdes et al in 2008 and was therefore used by team NUS Singapore 2019 as part of their sleep-wake module to control the growth of E. coli by inducing dormancy in these cells. it is used to demonstrate that when cells are sleeping, there is little luminescence production and when cells as awake, luminescence production resumes.
Biology
HicA originates from the hicAB locus of Escherichia coli K-12. HicA toxins cleave mRNAs independently of the ribosome. Overexpression leads to cleavage of a number of mRNAs and tmRNA, in a translation-independent fashion, suggesting that HicA is an mRNA interferase, which may play a role in bacterial resistance to antibiotics. In addition, overexpression of HicA leads to cell death and inhibits cell proliferation via inhibition of translation. The effect may be overcome by expression of antitoxin HicB.
References
Zhang, Y., Zhang, J., Hoeflich, K.P., Ikura, M., Qing, G. and Inouye, M. (2003) MazF cleaves cellular mRNAs specifically at ACA to block protein synthesis in Escherichia coli. Molecular Cell, 12, 913–923.
Jorgensen, M. G., Pandey, D. P., Jaskolska, M., & Gerdes, K. (2008). HicA of Escherichia coli Defines a Novel Family of Translation-Independent mRNA Interferases in Bacteria and Archaea. Journal of Bacteriology, 191(4), 1191–1199. doi: 10.1128/jb.01013-08
Maisonneuve, E., Shakespeare, L. J., Jørgensen, M. G., & Gerdes, K. (2011). Bacterial persistence by RNA endonucleases. Proceedings of the National Academy of Sciences, 108(32), 13206–13211. doi: 10.1073/pnas.1100186108
Source
BBa_K3198000 originated from E. coli K12 and its sequence was synthesized by IDT.
LuxABCDE is from Part:BBa_K325909.
Design Considerations
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 549
Illegal NheI site found at 1767
Illegal NheI site found at 4733 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 3731
Illegal BamHI site found at 489
Illegal BamHI site found at 1706
Illegal XhoI site found at 4561 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1541
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 6120
Illegal BsaI.rc site found at 3129
Illegal SapI site found at 1523
Illegal SapI.rc site found at 6445
| None |