1887

Microbiology Society logo

Volume 158, Issue 10

The P1 promoter is essential for the coordination of cell cycle events that prevent the overinitiation of DNA replication Free

Abstract

The master regulator CtrA oscillates during the cell cycle due to temporally regulated proteolysis and transcription. It is proteolysed during the G1–S transition and reaccumulates in predivisional cells as a result of transcription from two sequentially activated promoters, P1 and P2. CtrA reinforces its own synthesis by directly mediating the activation of P2 concurrently with repression of P1. To explore the role of P1 in cell cycle control, we engineered a mutation into the native locus that prevents transcription from P1 but not P2. As expected, the P1 mutant exhibits striking growth, morphological and DNA replication defects. Unexpectedly, we found CtrA and its antagonist SciP, but not DnaA, GcrA or CcrM accumulation to be dramatically reduced in the P1 mutant. SciP levels closely paralleled CtrA accumulation, suggesting that CtrA acts as a rheostat to modulate SciP abundance. Furthermore, the reappearance of CtrA and CcrM in predivisional cells was delayed in the P1 mutant by 0.125 cell cycle unit in synchronized cultures. High levels of transcription despite low levels of CtrA and increased transcription of P2 in the P1 mutant are two examples of robustness in the cell cycle. Thus, can adjust regulatory pathways to partially compensate for reduced and delayed CtrA accumulation in the P1 mutant.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
© Microbiology Society
Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.055285-0
2012-10-01
2024-04-20
Loading full text...

Full text loading...

/deliver/fulltext/micro/158/10/2492.html?itemId=/content/journal/micro/10.1099/mic.0.055285-0&mimeType=html&fmt=ahah

References

  1. Anderson P. E., Gober J. W. ( 2000). FlbT, the post-transcriptional regulator of flagellin synthesis in Caulobacter crescentus, interacts with the 5′ untranslated region of flagellin mRNA. Mol Microbiol 38:41–52 [View Article][PubMed]
     [Google Scholar]
  2. Argueta C., Yuksek K., Patel R., Summers M. L. ( 2006). Identification of Nostoc punctiforme akinete-expressed genes using differential display. Mol Microbiol 61:748–757 [View Article][PubMed]
     [Google Scholar]
  3. Ausmees N., Jacobs-Wagner C. ( 2003). Spatial and temporal control of differentiation and cell cycle progression in Caulobacter crescentus . Annu Rev Microbiol 57:225–247 [View Article][PubMed]
     [Google Scholar]
  4. Barboriak D. P., Padua A. O., York G. E., Macfall J. R. ( 2005). Creation of DICOM–aware applications using ImageJ. J Digit Imaging 18:91–99 [View Article][PubMed]
     [Google Scholar]
  5. Bastedo D. P., Marczynski G. T. ( 2009). CtrA response regulator binding to the Caulobacter chromosome replication origin is required during nutrient and antibiotic stress as well as during cell cycle progression. Mol Microbiol 72:139–154 [View Article][PubMed]
     [Google Scholar]
  6. Chen J. C., Hottes A. K., McAdams H. H., McGrath P. T., Viollier P. H., Shapiro L. ( 2006). Cytokinesis signals truncation of the PodJ polarity factor by a cell cycle-regulated protease. EMBO J 25:377–386 [View Article][PubMed]
     [Google Scholar]
  7. Christen B., Abeliuk E., Collier J. M., Kalogeraki V. S., Passarelli B., Coller J. A., Fero M. J., McAdams H. H., Shapiro L. ( 2011). The essential genome of a bacterium. Mol Syst Biol 7:528 [View Article][PubMed]
     [Google Scholar]
  8. Collier J., Shapiro L. ( 2009). Feedback control of DnaA-mediated replication initiation by replisome-associated HdaA protein in Caulobacter . J Bacteriol 191:5706–5716 [View Article][PubMed]
     [Google Scholar]
  9. Collier J., Murray S. R., Shapiro L. ( 2006). DnaA couples DNA replication and the expression of two cell cycle master regulators. EMBO J 25:346–356 [View Article][PubMed]
     [Google Scholar]
  10. Collier J., McAdams H. H., Shapiro L. ( 2007). A DNA methylation ratchet governs progression through a bacterial cell cycle. Proc Natl Acad Sci U S A 104:17111–17116 [View Article][PubMed]
     [Google Scholar]
  11. Cuajungco M. P., Podevin W., Valluri V. K., Bui Q., Nguyen V. H., Taylor K. ( 2012). Abnormal accumulation of human transmembrane (TMEM)-176A and 176B proteins is associated with cancer pathology. Acta Histochem 114:705–712 [View Article][PubMed]
     [Google Scholar]
  12. Domian I. J., Quon K. C., Shapiro L. ( 1997). Cell type-specific phosphorylation and proteolysis of a transcriptional regulator controls the G1-to-S transition in a bacterial cell cycle. Cell 90:415–424 [View Article][PubMed]
     [Google Scholar]
  13. Domian I. J., Reisenauer A., Shapiro L. ( 1999). Feedback control of a master bacterial cell-cycle regulator. Proc Natl Acad Sci U S A 96:6648–6653 [View Article][PubMed]
     [Google Scholar]
  14. Ely B. ( 1991). Genetics of Caulobacter crescentus . Methods Enzymol 204:372–384 [View Article][PubMed]
     [Google Scholar]
  15. Evinger M., Agabian N. ( 1977). Envelope-associated nucleoid from Caulobacter crescentus stalked and swarmer cells. J Bacteriol 132:294–301[PubMed]
     [Google Scholar]
  16. Gora K. G., Tsokos C. G., Chen Y. E., Srinivasan B. S., Perchuk B. S., Laub M. T. ( 2010). A cell-type-specific protein-protein interaction modulates transcriptional activity of a master regulator in Caulobacter crescentus . Mol Cell 39:455–467 [View Article][PubMed]
     [Google Scholar]
  17. Holtzendorff J., Hung D., Brende P., Reisenauer A., Viollier P. H., McAdams H. H., Shapiro L. ( 2004). Oscillating global regulators control the genetic circuit driving a bacterial cell cycle. Science 304:983–987 [View Article][PubMed]
     [Google Scholar]
  18. Hottes A. K., Shapiro L., McAdams H. H. ( 2005). DnaA coordinates replication initiation and cell cycle transcription in Caulobacter crescentus . Mol Microbiol 58:1340–1353 [View Article][PubMed]
     [Google Scholar]
  19. Iniesta A. A., McGrath P. T., Reisenauer A., McAdams H. H., Shapiro L. ( 2006). A phospho-signaling pathway controls the localization and activity of a protease complex critical for bacterial cell cycle progression. Proc Natl Acad Sci U S A 103:10935–10940 [View Article][PubMed]
     [Google Scholar]
  20. Jenal U., Fuchs T. ( 1998). An essential protease involved in bacterial cell-cycle control. EMBO J 17:5658–5669 [View Article][PubMed]
     [Google Scholar]
  21. Jonas K., Chen Y. E., Laub M. T. ( 2011). Modularity of the bacterial cell cycle enables independent spatial and temporal control of DNA replication. Curr Biol 21:1092–1101 [View Article][PubMed]
     [Google Scholar]
  22. Lam H., Schofield W. B., Jacobs-Wagner C. ( 2006). A landmark protein essential for establishing and perpetuating the polarity of a bacterial cell. Cell 124:1011–1023 [View Article][PubMed]
     [Google Scholar]
  23. Laub M. T., McAdams H. H., Feldblyum T., Fraser C. M., Shapiro L. ( 2000). Global analysis of the genetic network controlling a bacterial cell cycle. Science 290:2144–2148 [View Article][PubMed]
     [Google Scholar]
  24. Laub M. T., Chen S. L., Shapiro L., McAdams H. H. ( 2002). Genes directly controlled by CtrA, a master regulator of the Caulobacter cell cycle. Proc Natl Acad Sci U S A 99:4632–4637 [View Article][PubMed]
     [Google Scholar]
  25. Laub M. T., Shapiro L., McAdams H. H. ( 2007). Systems biology of Caulobacter . Annu Rev Genet 41:429–441 [View Article][PubMed]
     [Google Scholar]
  26. Lesley J. A., Shapiro L. ( 2008). SpoT regulates DnaA stability and initiation of DNA replication in carbon-starved Caulobacter crescentus . J Bacteriol 190:6867–6880 [View Article][PubMed]
     [Google Scholar]
  27. McAdams H. H., Shapiro L. ( 2011). The architecture and conservation pattern of whole-cell control circuitry. J Mol Biol 409:28–35 [View Article][PubMed]
     [Google Scholar]
  28. McGrath P. T., Iniesta A. A., Ryan K. R., Shapiro L., McAdams H. H. ( 2006). A dynamically localized protease complex and a polar specificity factor control a cell cycle master regulator. Cell 124:535–547 [View Article][PubMed]
     [Google Scholar]
  29. Meisenzahl A. C., Shapiro L., Jenal U. ( 1997). Isolation and characterization of a xylose-dependent promoter from Caulobacter crescentus . J Bacteriol 179:592–600[PubMed]
     [Google Scholar]
  30. Miller J. H. ( 1972). Experiments in Molecular Genetics Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
     [Google Scholar]
  31. Quon K. C., Marczynski G. T., Shapiro L. ( 1996). Cell cycle control by an essential bacterial two-component signal transduction protein. Cell 84:83–93 [View Article][PubMed]
     [Google Scholar]
  32. Quon K. C., Yang B., Domian I. J., Shapiro L., Marczynski G. T. ( 1998). Negative control of bacterial DNA replication by a cell cycle regulatory protein that binds at the chromosome origin. Proc Natl Acad Sci U S A 95:120–125 [View Article][PubMed]
     [Google Scholar]
  33. Reisenauer A., Shapiro L. ( 2002). DNA methylation affects the cell cycle transcription of the CtrA global regulator in Caulobacter . EMBO J 21:4969–4977 [View Article][PubMed]
     [Google Scholar]
  34. Reisenauer A., Quon K., Shapiro L. ( 1999). The CtrA response regulator mediates temporal control of gene expression during the Caulobacter cell cycle. J Bacteriol 181:2430–2439[PubMed]
     [Google Scholar]
  35. Sambrook J., Fritsch E. F., Maniatis T. ( 1989). Molecular Cloning: A Laboratory Manual Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
     [Google Scholar]
  36. Shen X., Collier J., Dill D., Shapiro L., Horowitz M., McAdams H. H. ( 2008). Architecture and inherent robustness of a bacterial cell-cycle control system. Proc Natl Acad Sci U S A 105:11340–11345 [View Article][PubMed]
     [Google Scholar]
  37. Simon R., Priefer U., Puhler A. ( 1983). A broad host range mobilization system for in vivo genetic engineering: transposon mutagenesis in Gram-negative bacteria. Biotechnology 1:784–791 [View Article]
     [Google Scholar]
  38. Spencer W., Siam R., Ouimet M. C., Bastedo D. P., Marczynski G. T. ( 2009). CtrA, a global response regulator, uses a distinct second category of weak DNA binding sites for cell cycle transcription control in Caulobacter crescentus . J Bacteriol 191:5458–5470 [View Article][PubMed]
     [Google Scholar]
  39. Stephens C., Reisenauer A., Wright R., Shapiro L. ( 1996). A cell cycle-regulated bacterial DNA methyltransferase is essential for viability. Proc Natl Acad Sci U S A 93:1210–1214 [View Article][PubMed]
     [Google Scholar]
  40. Tan M. H., Kozdon J. B., Shen X., Shapiro L., McAdams H. H. ( 2010). An essential transcription factor, SciP, enhances robustness of Caulobacter cell cycle regulation. Proc Natl Acad Sci U S A 107:18985–18990 [View Article][PubMed]
     [Google Scholar]
  41. Tsang P. H., Li G., Brun Y. V., Freund L. B., Tang J. X. ( 2006). Adhesion of single bacterial cells in the micronewton range. Proc Natl Acad Sci U S A 103:5764–5768 [View Article][PubMed]
     [Google Scholar]
  42. West L., Yang D., Stephens C. ( 2002). Use of the Caulobacter crescentus genome sequence to develop a method for systematic genetic mapping. J Bacteriol 184:2155–2166 [View Article][PubMed]
     [Google Scholar]
  43. Winzeler E., Shapiro L. ( 1995). Use of flow cytometry to identify a Caulobacter 4.5 S RNA temperature-sensitive mutant defective in the cell cycle. J Mol Biol 251:346–365 [View Article][PubMed]
     [Google Scholar]
  44. Zweiger G., Marczynski G., Shapiro L. ( 1994). A Caulobacter DNA methyltransferase that functions only in the predivisional cell. J Mol Biol 235:472–485 [View Article][PubMed]
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.055285-0
Loading
The Caulobacter crescentus ctrA P1 promoter is essential for the coordination of cell cycle events that prevent the overinitiation of DNA replication
Microbiology 158, 2492 (2012); https://doi.org/10.1099/mic.0.055285-0
/content/journal/micro/10.1099/mic.0.055285-0
/content/journal/micro/10.1099/mic.0.055285-0
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error