1887

Abstract

The presence and significance of developmentally regulated nucleases in ETH 7451 has been studied in relation to the lytic processes occurring during differentiation. The cell-death processes have been followed in surface cultures by a propidium iodide viability assay. This has allowed the visualization of dead (membrane-damaged, red fluorescent) and live (membrane-intact, green fluorescent) mycelium during development, and has facilitated the analysis of the role of nucleases in these processes. A parallel activity-gel analysis showed the appearance of 20–22 kDa, 34 kDa and 44 kDa nucleases, the latter appearing only when aerial mycelium is formed. The appearance of these nucleases shows a remarkable correlation with the death process of the mycelium during differentiation and with chromosomal DNA degradation. The 20–22 kDa enzymes are possibly related to the lytic phenomena taking place in the vegetative substrate mycelium before the emergence of the reproductive aerial mycelium, whereas the function of the 44 kDa nuclease seems to be related to the sporulation step. The 20–22 kDa nucleases require Ca for activity and are inhibited by Zn. The nucleases are loosely bound to the cell wall from where they can be liberated by simple washing. Conceivably, these enzymes work together and co-ordinate to achieve an efficient hydrolysis of DNA from dying cells. The results show that the biochemical reactions related with the lytic DNA degradation during the programmed cell death are notably conserved in . Some of the features of the process and the biochemical characteristics of the enzymes involved are analogous to those taking place during the DNA fragmentation processes in eukaryotic apoptotic cells.

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2002-02-01
2019-08-19
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References

  1. Aravind, L., Dixit, V. M. & Koonin, E. V. ( 1999; ). The domains of death: evolution of the apoptosis machinery. Trends Biochem Sci 24, 47-53.[CrossRef]
    [Google Scholar]
  2. Baltz, R. H. ( 1998; ). Genetic manipulation of antibiotic-producing Streptomyces. Trends Biochem Sci 6, 76-83.
    [Google Scholar]
  3. Cal, S., Aparicio, J. F., De los Reyes-Gavilán, C. G., Nicieza, R. G. & Sánchez, J. ( 1995; ). A novel exocytoplasmic endonuclease from Streptomyces antibioticus. Biochem J 306, 93-100.
    [Google Scholar]
  4. Chater, K. F. ( 1998; ). Taking a genetic scalpel to the Streptomyces colony. Microbiology 144, 1465-1478.[CrossRef]
    [Google Scholar]
  5. Fernández, M. & Sánchez, J. ( 2001; ). Viability staining and terminal deoxyribonucleotide transferase-mediated dUTP nick end labeling of the mycelium in submerged cultures of Streptomyces antibioticus ETH7451. J Microbiol Methods 47, 293-299.[CrossRef]
    [Google Scholar]
  6. Fernández, M., Olek, A., Walter, J. & Sánchez, J. ( 1998; ). Analysis of DNA methylation processes related to the inhibition of DNA synthesis by 5-azacytidine in Streptomyces antibioticus ETH7451. Biol Chem 379, 559-562.
    [Google Scholar]
  7. Granozzi, C., Billeta, R., Passantino, R., Sollazzo, M. & Puglia, A. M. ( 1990; ). A breakdown in macromolecular synthesis preceding differentiation in Streptomyces coelicolor A3(2). J Gen Microbiol 136, 713-716.[CrossRef]
    [Google Scholar]
  8. Hale, A. J., Smith, C. A., Sutherland, L. C., Stoneman, V. E. A., Longthorne, V. L., Culhane, A. C. & Williams, G. T. ( 1996; ). Apoptosis: molecular regulation of cell death. Eur J Biochem 236, 1-26.[CrossRef]
    [Google Scholar]
  9. Hochman, A. ( 1997; ). Programmed cell death in prokaryotes. Crit Rev Microbiol 23, 207-214.[CrossRef]
    [Google Scholar]
  10. Hughes, F. M. & Cidlowski, J. A. ( 1999; ). Potassium is a critical regulator of apoptotic enzymes in vitro and in vivo. Adv Enzyme Regul 39, 157-171.[CrossRef]
    [Google Scholar]
  11. Laemmli, U. K. ( 1970; ). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680-685.[CrossRef]
    [Google Scholar]
  12. Mendez, C., Braña, A. F., Manzanal, M. B. & Hardisson, C. ( 1985; ). Role of substrate mycelium in colony development in Streptomyces. Can J Microbiol 31, 446-450.[CrossRef]
    [Google Scholar]
  13. Miguelez, E. M., Hardisson, C. & Manzanal, M. B. ( 1999; ). Hyphal death during colony development in Streptomyces antibioticus: morphological evidence for the existence of a process of cell deletion equivalent to apoptosis in a multicellular prokaryote. J Cell Biol 145, 515-525.[CrossRef]
    [Google Scholar]
  14. Montague, J. W. & Cidlowski, J. A. ( 1996; ). Cellular catabolism in apoptosis: DNA degradation and endonuclease activation. Experientia 52, 957-962.[CrossRef]
    [Google Scholar]
  15. Nicieza, R. G., Huergo, J., Connolly, B. A. & Sánchez, J. ( 1999; ). Purification, characterization and role of nucleases and serine proteases in Streptomyces differentiation: analogies with the biochemical processes described in late-steps of eukaryotic apoptosis. J Biol Chem 274, 20366-20375.[CrossRef]
    [Google Scholar]
  16. Novella, I. S., Barbés, C. & Sánchez, J. ( 1992; ). Sporulation of Streptomyces antibioticus ETHZ 7451 in submerged culture. Can J Microbiol 38, 769-773.[CrossRef]
    [Google Scholar]
  17. Peitsch, M. C., Mannherz, X. G. & Tschopp, J. ( 1994; ). The apoptosis endonucleases: cleaning up after cell death? Trends Cell Biol 4, 37-41.[CrossRef]
    [Google Scholar]
  18. Rosenthal, A. & Lacks, S. A. ( 1977; ). Nuclease detection in SDS-polyacrylamide gel electrophoresis. Anal Biochem 80, 76-90.[CrossRef]
    [Google Scholar]
  19. Smith, R. J. ( 1995; ). Calcium and bacteria. Adv Microb Physiol 37, 83-103.
    [Google Scholar]
  20. Widlak, P. & Garrard, W. T. ( 2001; ). Ionic and cofactor requirements for the activity of the apoptotic endonuclease DFF40/CAD. Mol Cell Biochem 218, 125-130.[CrossRef]
    [Google Scholar]
  21. Yarmolinsky, M. B. ( 1995; ). Programmed cell death in bacterial populations. Science 267, 836-837.[CrossRef]
    [Google Scholar]
  22. Zhang, C.-C. ( 1996; ). Bacterial signalling involving eukaryotic-type protein kinases. Mol Microbiol 20, 9-15.[CrossRef]
    [Google Scholar]
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