Abstract
The different Escherichia coli envelope fractions (cell wall, cytoplasmic membrane, and DNA-envelope complex fragments) were isolated by free-flow electrophoresis and analyzed by sodium dodecylsulfate-acrylamide gel electrophoresis. The DNA-envelope complex fragments possess a specific protein (mol wt 80,000-90,000). Upon treatment with trypsin, this protein disappears and the complex breaks down, thus releasing DNA, cell wall, and cytoplasmic membrane. Disaggregation of the complex can also be achieved by high salt concentrations. Lysozyme treatment dissolves the murein layer within the complex but does not disaggregate the complex. From these and other results on the stability of the DNA-envelope complex, conclusions can be drawn about the possible linkage within the described envelope particles.
Full Text
The Full Text of this article is available as a PDF (4.0 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Ames G. F., Spudich E. N., Nikaido H. Protein composition of the outer membrane of Salmonella typhimurium: effect of lipopolysaccharide mutations. J Bacteriol. 1974 Feb;117(2):406–416. doi: 10.1128/jb.117.2.406-416.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bayer M. E. Areas of adhesion between wall and membrane of Escherichia coli. J Gen Microbiol. 1968 Oct;53(3):395–404. doi: 10.1099/00221287-53-3-395. [DOI] [PubMed] [Google Scholar]
- Braun V., Rehn K. Chemical characterization, spatial distribution and function of a lipoprotein (murein-lipoprotein) of the E. coli cell wall. The specific effect of trypsin on the membrane structure. Eur J Biochem. 1969 Oct;10(3):426–438. doi: 10.1111/j.1432-1033.1969.tb00707.x. [DOI] [PubMed] [Google Scholar]
- Braun V., Rehn K., Wolff H. Supramolecular structure of the rigid layer of the cell wall of Salmonella, Serratia, Proteus, and Pseudomonas fluorescens. Number of lipoprotein molecules in a membrane layer. Biochemistry. 1970 Dec 22;9(26):5041–5049. doi: 10.1021/bi00828a001. [DOI] [PubMed] [Google Scholar]
- Braun V., Sieglin U. The covalent murein-lipoprotein structure of the Escherichia coli cell wall. The attachment site of the lipoprotein on the murein. Eur J Biochem. 1970 Apr;13(2):336–346. doi: 10.1111/j.1432-1033.1970.tb00936.x. [DOI] [PubMed] [Google Scholar]
- Costerton J. W., Ingram J. M., Cheng K. J. Structure and function of the cell envelope of gram-negative bacteria. Bacteriol Rev. 1974 Mar;38(1):87–110. doi: 10.1128/br.38.1.87-110.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
- De Petris S. Ultrastructure of the cell wall of Escherichia coli and chemical nature of its constituent layers. J Ultrastruct Res. 1967 Jul;19(1):45–83. doi: 10.1016/s0022-5320(67)80059-5. [DOI] [PubMed] [Google Scholar]
- DePamphilis M. L., Adler J. Attachment of flagellar basal bodies to the cell envelope: specific attachment to the outer, lipopolysaccharide membrane and the cyoplasmic membrane. J Bacteriol. 1971 Jan;105(1):396–407. doi: 10.1128/jb.105.1.396-407.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dingman C. W. Bidirectional chromosome replication: some topological considerations. J Theor Biol. 1974 Jan;43(1):187–195. doi: 10.1016/s0022-5193(74)80052-4. [DOI] [PubMed] [Google Scholar]
- Earhart C. F., Tremblay G. Y., Daniels M. J., Schaechter M. DNA replication studied by a new method for the isolation of cell membrane-DNA complexes. Cold Spring Harb Symp Quant Biol. 1968;33:707–710. doi: 10.1101/sqb.1968.033.01.079. [DOI] [PubMed] [Google Scholar]
- Fairbanks G., Steck T. L., Wallach D. F. Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry. 1971 Jun 22;10(13):2606–2617. doi: 10.1021/bi00789a030. [DOI] [PubMed] [Google Scholar]
- Fielding P., Fox C. F. Evidence for stable attachment of DNA to membrane at the replication origin of Escherichia coli. Biochem Biophys Res Commun. 1970 Oct 9;41(1):157–162. doi: 10.1016/0006-291x(70)90482-1. [DOI] [PubMed] [Google Scholar]
- Fuchs E., Hanawalt P. Isolation and characterization of the DNA replication complex from Escherichia coli. J Mol Biol. 1970 Sep 14;52(2):301–322. doi: 10.1016/0022-2836(70)90032-x. [DOI] [PubMed] [Google Scholar]
- Glauert A. M., Thornley M. J. The topography of the bacterial cell wall. Annu Rev Microbiol. 1969;23:159–198. doi: 10.1146/annurev.mi.23.100169.001111. [DOI] [PubMed] [Google Scholar]
- HANAWALT P. C., RAY D. S. ISOLATION OF THE GROWING POINT IN THE BACTERIAL CHROMOSOME. Proc Natl Acad Sci U S A. 1964 Jul;52:125–132. doi: 10.1073/pnas.52.1.125. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hannig K., Heidrich H. G. The use of continuous preparative free-flow electrophoresis for dissociating cell fractions and isolation of membranous components. Methods Enzymol. 1974;31:746–761. doi: 10.1016/0076-6879(74)31078-6. [DOI] [PubMed] [Google Scholar]
- Heidrich H. G. New aspects on the heterogeneity of beef liver catalase. Hoppe Seylers Z Physiol Chem. 1968 Jul;349(7):873–880. doi: 10.1515/bchm2.1968.349.2.873. [DOI] [PubMed] [Google Scholar]
- Henning U., Höhn B., Sonntag I. Cell envelope and shape of Escherichia coli K12. The ghost membrane. Eur J Biochem. 1973 Nov 1;39(1):27–36. doi: 10.1111/j.1432-1033.1973.tb03099.x. [DOI] [PubMed] [Google Scholar]
- Holland I. B., Darby V. Detection of a specific DNA-cytoplasmic membrane complex in Escherichia coli by equilibrium density centrifugation on sucrose gradients. FEBS Lett. 1973 Jun 15;33(1):106–108. doi: 10.1016/0014-5793(73)80170-x. [DOI] [PubMed] [Google Scholar]
- Inouye M. A three-dimensional molecular assembly model of a lipoprotein from the Escherichia coli outer membrane. Proc Natl Acad Sci U S A. 1974 Jun;71(6):2396–2400. doi: 10.1073/pnas.71.6.2396. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Inouye M., Guthrie J. P. A mutation which changes a membrane protein of E. coli. Proc Natl Acad Sci U S A. 1969 Nov;64(3):957–961. doi: 10.1073/pnas.64.3.957. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Inouye M., Pardee A. B. Changes of membrane proteins and their relation to deoxyribonucleic acid synthesis and cell division of Escherichia coli. J Biol Chem. 1970 Nov 10;245(21):5813–5819. [PubMed] [Google Scholar]
- Inouye M., Yee M. L. Homogeneity of envelope proteins of Escherichia coli separated by gel electrophoresis in sodium dodecyl sulfate. J Bacteriol. 1973 Jan;113(1):304–312. doi: 10.1128/jb.113.1.304-312.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Klein A., Bonhoeffer F. DNA replication. Annu Rev Biochem. 1972;41(10):301–332. doi: 10.1146/annurev.bi.41.070172.001505. [DOI] [PubMed] [Google Scholar]
- Lazdunski A., Shapiro B. M. The significance of membrane alterations seen in DNA synthesis mutants of Escherichia coli. Biochim Biophys Acta. 1973 Feb 27;298(1):59–68. doi: 10.1016/0005-2736(73)90009-6. [DOI] [PubMed] [Google Scholar]
- Olsen W. L., Heidrich H. G., Hannig K., Hofschneider P. H. Deoxyribonucleic acid-envelope complexes isolated from Escherichia coli by free-flow electrophoresis: biochemical and electron microscope characterization. J Bacteriol. 1974 May;118(2):646–653. doi: 10.1128/jb.118.2.646-653.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Osborn M. J., Gander J. E., Parisi E., Carson J. Mechanism of assembly of the outer membrane of Salmonella typhimurium. Isolation and characterization of cytoplasmic and outer membrane. J Biol Chem. 1972 Jun 25;247(12):3962–3972. [PubMed] [Google Scholar]
- Rice T. W., Grula E. A. Effects of temperature on extraction and electrophoretic migration of envelope proteins from Erwinia species. Biochim Biophys Acta. 1974 Mar 14;342(1):125–132. doi: 10.1016/0005-2795(74)90113-5. [DOI] [PubMed] [Google Scholar]
- Ryter A. Association of the nucleus and the membrane of bacteria: a morphological study. Bacteriol Rev. 1968 Mar;32(1):39–54. doi: 10.1128/br.32.1.39-54.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schnaitman C. A. Effect of ethylenediaminetetraacetic acid, Triton X-100, and lysozyme on the morphology and chemical composition of isolate cell walls of Escherichia coli. J Bacteriol. 1971 Oct;108(1):553–563. doi: 10.1128/jb.108.1.553-563.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schnaitman C. A. Solubilization of the cytoplasmic membrane of Escherichia coli by Triton X-100. J Bacteriol. 1971 Oct;108(1):545–552. doi: 10.1128/jb.108.1.545-552.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shapiro B. M., Siccardi A. G., Hirota Y., Jacob F. On the process of cellular division in Escherichia coli. Membrane prtein alterations associated with mutations affecting the initiation of DNA synthesis. J Mol Biol. 1970 Aug 28;52(1):75–89. doi: 10.1016/0022-2836(70)90178-6. [DOI] [PubMed] [Google Scholar]
- Siegel P. J., Schaechter M. The role of the host cell membrane in the replication and morphogenesis of bacteriophages. Annu Rev Microbiol. 1973;27:261–282. doi: 10.1146/annurev.mi.27.100173.001401. [DOI] [PubMed] [Google Scholar]
- Smith D. W., Hanawalt P. C. Properties of the growing point region in the bacterial chromosome. Biochim Biophys Acta. 1967 Dec 19;149(2):519–531. doi: 10.1016/0005-2787(67)90180-3. [DOI] [PubMed] [Google Scholar]
- Spencer M. E., Guest J. R. Proteins of the inner membrane of Escherichia coli: identification of succinate dehydrogenase by polyacrylamide gel electrophoresis with sdh amber mutants. J Bacteriol. 1974 Mar;117(3):947–953. doi: 10.1128/jb.117.3.947-953.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sueoka N., Hammers J. M. Isolation of DNA-membrane complex in Bacillus subtilis. Proc Natl Acad Sci U S A. 1974 Dec;71(12):4787–4791. doi: 10.1073/pnas.71.12.4787. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sueoka N., Quinn W. G. Membrane attachment of the chromosome replication origin in Bacillus subtilis. Cold Spring Harb Symp Quant Biol. 1968;33:695–705. doi: 10.1101/sqb.1968.033.01.078. [DOI] [PubMed] [Google Scholar]
- Tremblay G. Y., Daniels M. J., Schaechter M. Isolation of a cell membrane-DNA-nascent RNA complex from bacteria. J Mol Biol. 1969 Feb 28;40(1):65–76. doi: 10.1016/0022-2836(69)90296-4. [DOI] [PubMed] [Google Scholar]