Abstract
Intracellular catabolism of proteins labeled at the end of the exponential growth proceeded in two phases during sporulation. The first phase was induced by starvation and took place also in cells whose sporulation was inhibited by netropsin. The second phase of degradation, which was triggered at the onset of the irreversible sporulation phase, was inhibited by netropsin. Intracellular proteolytic activity determined in disintegrated cells, i.e., primarily the activity of the cytoplasmic Ca2+-dependent serine proteinase(s) at the first place, was increasing throughout the sporulation process and reached its maximum during the irreversible sporulation phase. Its increase was suppressed by netropsin. Fractionation of the cell sap by HPLC revealed a similar distribution of proteolytic activities in the extract from control and netropsin-inhibited cells. The antibiotic thus probably affected the activation, not the formation of the cytoplasmic serine proteinase(s). Netropsin also inhibited an increase of proteolytic activity in the membrane fraction, probably owing to the presence of two different proteolytic enzymes.
Similar content being viewed by others
Literature Cited
Band J, Henner DJ, Ruppen M (1987) Construction and properties of an intracellular proteinase mutant ofBacillus subtilis. J Bacteriol 169:444–446
Burnett TJ, Shankweiler GW, Hageman JH (1986) Activation of intracellular serine proteinase inBacillus subtilis cells during sporulation. J Bacteriol 165:139–145
Chaloupka J, Kučerová H (1988) Netropsin increases formation of mRNA coding for a neutral metalloproteinase inBacillus megaterium. J Basic Microbiol 28:11–16
Chaloupka J, Severin AI, Sastry KJ, Kučerová H, Strnadová M (1982) Differences in the regulation of exocellular proteinase synthesis during growth and sporogenesis ofBacillus megaterium. Can J Microbiol 28:214–218
Cheng YSE, Aronson AI (1977a) Characterization and function of intracellular proteases in sporulatingBacillus cereus. Arch Microbiol 115:61–66
Cheng YSE, Aronson AI (1977b) Alteration of spore coat processing and protein turnover in aBacillus cereus mutant with a defective postexponential intracellular protease. Proc Natl Acad Sci USA 74:1254–1258
Guinand M, Vacheron MJ, Michel G (1978) Relation between inhibition of bacilli sporulation and synthesis of lytic enzymes. Biochem Biophys Res Commun 80:429–434
Hageman JH, Carlton BC (1973) Effect of mutational loss of specific intracellular proteases on the sporulation ofBacillus subtilis. J Bacteriol 114:612–617
Käs E, Izaurralde E, Laemmli UK (1989) Specific inhibition of DNA binding to nuclear scaffolds and histone H1 by distamycin. The role of oligo (dA). oligo (dT) tracts. J Mol Biol 210:587–599
Keilman GR, Tanimoto B, Doi RH (1975) Selective inhibition of sporulation ofBacillus subtilis by netropsin. Biochem Biophys Res Commun 67:414–420
Keilman GR, Burtis K, Tanimoto B, Doi RH (1976) Effect of netropsin on the derepression of enzymes during growth and sporulation ofBacillus subtilis. J Bacteriol 128:80–85
Kerjan P, Keryer E, Szulmajster J (1979) Characterization of a thermosensitive sporulation mutant ofBacillus subtilis affected in the structural gene of an intracellular protease. Eur J Biochem 98:353–362
Kopka ML, Yoon C, Goodsell D, Pjura P, Dickerson RE (1985) The molecular origin of DNA-drug specificity in netropsin and distamycin. Proc Natl Acad Sci USA 82:1376–1380
Kučerová H, Chaloupka J (1985) Suppression of an extracellular metalloproteinase synthesis inBacillus megaterium by temperature. FEMS Microbiol Lett 28:293–296
Kučerová H, Váchová L, Chaloupka J (1984) Mutants ofBacillus megaterium with altered synthesis of an extracellular proteinase. Folia Microbiol 29:99–103
Kučerová H, Strnadová M, Vinter V, Graba P, Chaloupka J (1986) Netropsin stimulates the formation of an extracellular proteinase and suppresses protein turnover in sporulatingBacillus megaterium. FEMS Microbiol Lett 34:21–26
Lavigne M, Herbert M, Kolb A, Buc H (1992) Upstream curved sequences influence the initiation of transcription at theEscherichia coli galactose operon. J Mol Biol 224:293–306
Millet J (1971) Caractérisation d'une endopeptidase cytoplasmic chezBacillus megaterium en voie de sporulation. Compt Rend Acad Sci 272:1806–1809
Millet J (1977) Characterization of a protein inhibitor of intracellular protease fromBacillus subtilis. FEBS Lett. 74:59–61
Moravcová J, Chaloupka J (1990) Characteristics of intracellular proteolytic activities ofBacillus megaterium. Folia Microbiol (Prague) 35:402–412
Munoz L. Sadaie Y, Doi RH (1978) Spore coat protein ofBacillus subtilis. Structure and precursor synthesis. J Biol Chem 253:6694–6701
Reysset G, Millet J (1972) Characterization of an intracellular protease inBacillus subtilis during sporulation. Biochem Biophys Res Commun 40:328–334
Ruppen ME, Van Alstine GL, Band L (1988) Control of intracellular serine protease expression inBacillus subtilis. J Bacteriol 170:136–140
Sheehan SM, Switzer RL (1991) Intracellular protease-4, a new intracellular serine protease activity fromBacillus subtilis. Arch Microbiol 156:186–191
Strongin AY, Izotova LS, Abramov ZT, Ermakova LM, Gorodetsky DI, Stepanov MV (1978) On the appearance ofBacillus subtilis intracellular serine proteinase in the cell membrane and culture medium. Comparison of the enzyme with otherBacillus subtilis serine proteinases. Arch Microbiol 119:287–293
Votruba J, Pazlarová J, Dvořáková M, Váchová L, Strnadová M, Kučerová H, Vinter V, Zourabian R, Chaloupka J (1991) External factors involved in the regulation of synthesis of an extracellular proteinase inBacillus megaterium: effect of temperature. Appl Microbiol Biotechnol 35:352–357
Wartell RM, Larson JE, Wells RD (1974) Netropsin. A specific probe for A-T regions of duplex deoxyribonucleic acid. J Biol Chem 249:6719–6731
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Kučerová, H., Moravcová, J., Váchová, L. et al. Netropsin suppresses the rise of activity of an intracellular proteolytic system in sporulatingBacillus megaterium . Current Microbiology 26, 287–292 (1993). https://doi.org/10.1007/BF01575919
Issue Date:
DOI: https://doi.org/10.1007/BF01575919