Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1980 Nov;144(2):789–799. doi: 10.1128/jb.144.2.789-799.1980

Bacillus megaterium sporal peptidoglycan synthesis studied by high-resolution autoradiography.

C Frehel, A Ryter
PMCID: PMC294730  PMID: 6776097

Abstract

Cells of a Dap- Lys- mutant strain of Bacillus megaterium were pulse labeled with [3H]diaminopimelic acid at different times of growth and sporulation. They were processed for radioactivity measurements and high-resolution autoradiography either just after the pulse or after a chase in a nonradioactive medium until refractile forespores started to appear at time (t)4,5. In the pulse-labeled cells, autoradiographs and radioactivity measurements showed that the radioactivity incorporated during a pulse decreased abruptly after t0 and stayed at a low level until t5, although the forespore wall and cortex were formed between t4 and t5. In the pulse-chased bacteria, the acid-insoluble radioactivity, as well as the number of silver grains on autoradiographs, increased during the chase in cells labeled at t1 to t2, whereas it decreased in those labeled before t0. Furthermore, analysis of silver grain distribution showed that, in stage IV bacteria, grains were distributed at the outside of the forespore, mostly on the sporangium cell wall, when pulse-labeling occurred before or at t0; they were located along the cortex and in the forespore cytoplasm when labeling was made at t1 or t2. These facts show that [3H]diaminopimelic acid necessary for spore envelope synthesis was incorporated before their morphological appearance. Free or small diaminopimelic acid precursors entered the sporangium between t1 and t2. The appearance of silver grains in the forespore cytoplasm suggests that the forespore is implicated in sporal peptidoglycan synthesis.

Full text

PDF
789

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aronson A. I., Fitz-James P. C. Biosynthesis of bacterial spore coats. J Mol Biol. 1968 Apr 14;33(1):199–212. doi: 10.1016/0022-2836(68)90288-x. [DOI] [PubMed] [Google Scholar]
  2. Baillie E., Murrell W. G. Some hydrodynamic properties of spore peptidoglycan. Biochim Biophys Acta. 1974 Nov 4;372(1):23–31. doi: 10.1016/0304-4165(74)90069-5. [DOI] [PubMed] [Google Scholar]
  3. Caro L. Progress in high-resolution autoradiography. Prog Biophys Mol Biol. 1966;16:171–190. doi: 10.1016/0079-6107(66)90006-x. [DOI] [PubMed] [Google Scholar]
  4. Coote J. G., Mandelstam J. Use of constructed double mutants for determining the temporal order of expression of sporulation genes in Bacillus subtilis. J Bacteriol. 1973 Jun;114(3):1254–1263. doi: 10.1128/jb.114.3.1254-1263.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Eaton M. W., Ellar D. J. Protein synthesis and breakdown in the mother-cell and forespore compartments during spore morphogenesis in Bacillus megaterium. Biochem J. 1974 Nov;144(2):327–337. doi: 10.1042/bj1440327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Frehel C., Ryter A. Peptidoglycan turnover during growth of a Bacillus megaterium Dap- Lys- mutant. J Bacteriol. 1979 Feb;137(2):947–955. doi: 10.1128/jb.137.2.947-955.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fréhel C., Ryter A. Réversibilité de la sporulation chez B. subtilis. Ann Inst Pasteur (Paris) 1969 Sep;117(3):297–311. [PubMed] [Google Scholar]
  8. Guinand M., Vacheron M. J., Michel G., Tipper D. J. Location of peptidoglycan lytic enzymes in Bacillus sphaericus. J Bacteriol. 1979 Apr;138(1):126–132. doi: 10.1128/jb.138.1.126-132.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Holt S. C., Gauther J. J., Tipper D. J. Ultrastructural studies of sporulation in Bacillus sphaericus. J Bacteriol. 1975 Jun;122(3):1322–1338. doi: 10.1128/jb.122.3.1322-1338.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. LUFT J. H. Improvements in epoxy resin embedding methods. J Biophys Biochem Cytol. 1961 Feb;9:409–414. doi: 10.1083/jcb.9.2.409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Linnett P. E., Tipper D. J. Transcriptional control of peptidoglycan precursor synthesis during sporulation in Bacillus sphaericus. J Bacteriol. 1976 Feb;125(2):565–574. doi: 10.1128/jb.125.2.565-574.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Pearce S. M. An invitro system for the biosynthesis of spore cortex peptidoglycan. Can J Microbiol. 1977 Aug;23(8):964–974. doi: 10.1139/m77-144. [DOI] [PubMed] [Google Scholar]
  13. Pitel D. W., Gilvarg C. Mucopeptide metabolism during growth and sporulation in Bacillus megaterium. J Biol Chem. 1970 Dec 25;245(24):6711–6717. [PubMed] [Google Scholar]
  14. RYTER A., KELLENBERGER E., BIRCHANDERSEN A., MAALOE O. Etude au microscope électronique de plasmas contenant de l'acide désoxyribonucliéique. I. Les nucléoides des bactéries en croissance active. Z Naturforsch B. 1958 Sep;13B(9):597–605. [PubMed] [Google Scholar]
  15. Ryter A., Whitehouse R. Uracil incorporation in the forespore and the mother cell during spore development in Bacillus subtilis. Autoradiographic electron microscopic study. Arch Microbiol. 1978 Jul;118(1):27–34. doi: 10.1007/BF00406070. [DOI] [PubMed] [Google Scholar]
  16. Salpeter M. M., Bachmann L., Salpeter E. E. Resolution in electron microscope radioautography. J Cell Biol. 1969 Apr;41(1):1–32. doi: 10.1083/jcb.41.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Singh R. P., Setlow B., Setlow P. Levels of small molecules and enzymes in the mother cell compartment and the forespore of sporulating Bacillus megaterium. J Bacteriol. 1977 Jun;130(3):1130–1138. doi: 10.1128/jb.130.3.1130-1138.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Tipper D. J., Linnett P. E. Distribution of peptidoglycan synthetase activities between sporangia and forespores in sporulating cells of Bacillus sphaericus. J Bacteriol. 1976 Apr;126(1):213–221. doi: 10.1128/jb.126.1.213-221.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. WARTH A. D., OHYE D. F., MURRELL W. G. Location and composition of spore mucopeptide in Bacillus species. J Cell Biol. 1963 Mar;16:593–609. doi: 10.1083/jcb.16.3.593. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Warth A. D., Strominger J. L. Structure of the peptidoglycan from spores of Bacillus subtilis. Biochemistry. 1972 Apr 11;11(8):1389–1396. doi: 10.1021/bi00758a010. [DOI] [PubMed] [Google Scholar]
  21. de Chastellier C., Frehel C., Ryter A. Cell wall growth of Bacillus megaterium: cytoplasmic radioactivity after pulse-labeling with tritiated diaminopimelic acid. J Bacteriol. 1975 Sep;123(3):1197–1207. doi: 10.1128/jb.123.3.1197-1207.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. de Chastellier C., Hellio R., Ryter A. Study of cell wall growth in Bacillus megaterium by high-resolution autoradiography. J Bacteriol. 1975 Sep;123(3):1184–1196. doi: 10.1128/jb.123.3.1184-1196.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES