Part:BBa_K2588005

DNA Replication inhibitor cspD

The cold shock protein D (cspD) is a replication inhibitor found in E. coli.

Usage and Biology

CspD was used as a part of the growth inhibition module. It is a small protein found in E. coli, which forms homodimers. It is induced in the stationary phase or upon carbon starvation and is inversely correlated with growth rate. Furthermore, it is involved in persister cell forming.^1,2
CspD forms β-barrels with two nucleic acid binding motifs to bind ssDNA and RNA there for no specific nucleotide sequence is needed. During replication cspD binds ssDNA mostly at replication forks also cspD inhibits the elongations steps. The Lon protease degrades the unstable cspD protein.^3,4
We cloned a RBS (BBa_B00XX) upstream of the cspD gene to overexpress it and so to stop the growth of E. coli. Luckily the expression of cspD is not induced by cold shock so we can use this part in the heat of malaria regions.⁵

cspD was used in our growth inhibition module, BBa_K2588021.

References

1. Yamanaka, K., Zheng, W., Crooke, E., Wang, Y.-H. & Inouye, M. CspD, a novel DNA replication inhibitor induced during the stationary phase in Escherichia coli. Mol. Microbiol. 39, 1572–1584 (2001).
2. Kim, Y. & Wood, T. K. Toxins Hha and CspD and small RNA regulator Hfq are involved in persister cell formation through MqsR in Escherichia coli. Biochem. Biophys. Res. Commun. 391, 209–213 (2010).
3. Langklotz, S. & Narberhaus, F. The Escherichia coli replication inhibitor CspD is subject to growth-regulated degradation by the Lon protease. Mol. Microbiol. 80, 1313–1325 (2011).
4. Yamanaka, K. & Inouye, M. Growth-phase-dependent expression of cspD, encoding a member of the CspA family in Escherichia coli. J. Bacteriol. 179, 5126–30 (1997).
5. Lee, S. J. et al. Family of the major cold-shock protein, CspA (CS7.4), of Escherichia coli, whose members show a high sequence similarity with the eukaryotic Y-box binding proteins. Mol. Microbiol. 11, 833–839 (1994).

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]