From 2010.igem.org
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Filamentous Cells
Filamentous cells genes list
- yneA (Transcribed with yneB, ynzC, is an analogue of sulA in E.coli) * our biobrick is designed to over express this gene reducing cell division possibly by inhibiting FtsZ ring formation or constriction.
- dinR (Homologue of lexA in E.coli transcribed in the opposite direction)
- ftsZ (Involved in the recruitment of other proteins to the divisisome for cytokinesis, strangely over expression results in disruption of Zring formation as well as reduced expression)
- secA (Involved in the secretion of extracellular proteins and the insertion of transmembrane proteins)
- recA (Involved in SOS response removing the repressor DinR (LexA))
- wpr and epr produce extracellular proteases that cleave the signal peptide/transmembrane domain of YneA
- ezr produces a protein which sequesters FtsZ monomer by binding its C terminal domain and also inhibits GTP binding; however overexpression does not result in filamentation.
- min C,D ,J and divIVA prevent polar cell division .
- Positive regulators of FtsZ: ftsA, zapA, zipA, ftsL and divIC
- Inhibitors of Daughter cell separation: lytC,D,E,F and cwlS *Chains rather than filaments, yneA is also reported to increase the time spent in chains well into the stationary phase of bacterial growth.
YneA
Kawai, Y., Moriya, S., & Ogasawara, N. (2003). Identification of a protein, YneA, responsible for cell division suppression during the SOS response in Bacillus subtilis. Molecular microbiology, 47(4), 1113-22. Retrieved from http://www.ncbi.nlm.nih.gov/pubmed/12581363.
yneA cloning strategy
Media:yneA cloning strategy.pdf
MinCDJ
In B.subtilis cell division occurs precisely mid cell via the formation of the FtsZ ring (tubulin homologue), through Noc (Nucleoid occlusion: prevents division over nucleoids) and the min system (well described in E.coli), which prevents division taking place at the ends of the cell (poles).
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Main function of Min system to prevent mini cell formation and ensuring only one cell division occurs per cell cycle.
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Min’s role is not just in the inhibition of FtsZ. FtsZ recruits other components causing synthesis of the new wall and cell invagination.
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![Min2.png](/wiki/images/b/b6/Min2.png)
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Cell division is regulated spatially and temporally.
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Min C inhibits FtsZ ring formation; Min C interacts with Min D via its C-terminal domain. Min C inhibits lateral interaction between the filaments.
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Min D is a membrane associated ATPase. (Min E ensures high concentrations of MinCD at the poles in E.coli, no Min E homologue in B.subtilis! Instead DivIVA acts as a topological factor).
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Min C in B.subtilis is responsible for localisation (shown using GFP) showed a primary site of localisation at the site of active division challenging the original model for the role of Min.
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Min J was discovered linking Min D to DivIVA and therefore necessary for localisation.
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Over expression of Min D in the absence of Min J causes lethal filamentation.
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Min C sequence
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http://www.ncbi.nlm.nih.gov/nuccore/NC_000964.3?from=2858550&to=2859298&strand=true&report=graph&content=5
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http://www.ncbi.nlm.nih.gov/gene/937500
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MinD sequence
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http://www.ncbi.nlm.nih.gov/nuccore/NC_000964.3?from=2857736&to=2858622&strand=true&report=graph&content=5
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http://www.ncbi.nlm.nih.gov/gene/937499
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MinJ sequence
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http://www.ncbi.nlm.nih.gov/nuccore/NC_000964.3?from=3620287&to=3621598&strand=true&report=graph&content=5
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http://www.ncbi.nlm.nih.gov/gene/936668
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van Baarle, S., & Bramkamp, M. (2010). The MinCDJ system in Bacillus subtilis prevents minicell formation by promoting divisome disassembly. PloS one, 5(3), e9850. doi: 10.1371/journal.pone.0009850.
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Bramkamp, M. & van Baarle, S., 2009. Division site selection in rod-shaped bacteria. Current opinion in microbiology, 12(6), 683-8. Available at: http://www.ncbi.nlm.nih.gov/pubmed/19884039.
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