Abstract
It has previously been shown that hemimethylated DNA from the Escherichia coli replication origin (oriC) binds with high specificity to membrane fractions isolated from disrupted cells. In this article, the membrane localization of oriC-binding activity was studied by subjecting crude membrane preparations to successive cycles of sedimentation and flotation gradient analysis. This revealed that approximately two-thirds of the membrane-associated oriC-binding activity of the cell was not associated with the outer membrane fraction as previously suggested but was recovered instead in a unique membrane fraction (OCB1) whose buoyant density and protein profile differed from those of both inner and outer membranes. The specific activity of oriC binding in OCB1 was approximately fivefold higher than the activity of the isolated outer membrane peak. It is likely that membrane fraction OCB1 includes the membrane domain responsible for the binding of hemimethylated oriC to the cell envelope in intact cells.
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- Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
- Buhk H. J., Messer W. The replication origin region of Escherichia coli: nucleotide sequence and functional units. Gene. 1983 Oct;24(2-3):265–279. doi: 10.1016/0378-1119(83)90087-2. [DOI] [PubMed] [Google Scholar]
- Casaregola S., Norris V., Goldberg M., Holland I. B. Identification of a 180 kD protein in Escherichia coli related to a yeast heavy-chain myosin. Mol Microbiol. 1990 Mar;4(3):505–511. doi: 10.1111/j.1365-2958.1990.tb00617.x. [DOI] [PubMed] [Google Scholar]
- Hendrickson W. G., Kusano T., Yamaki H., Balakrishnan R., King M., Murchie J., Schaechter M. Binding of the origin of replication of Escherichia coli to the outer membrane. Cell. 1982 Oct;30(3):915–923. doi: 10.1016/0092-8674(82)90296-3. [DOI] [PubMed] [Google Scholar]
- Ishidate K., Creeger E. S., Zrike J., Deb S., Glauner B., MacAlister T. J., Rothfield L. I. Isolation of differentiated membrane domains from Escherichia coli and Salmonella typhimurium, including a fraction containing attachment sites between the inner and outer membranes and the murein skeleton of the cell envelope. J Biol Chem. 1986 Jan 5;261(1):428–443. [PubMed] [Google Scholar]
- Kusano T., Steinmetz D., Hendrickson W. G., Murchie J., King M., Benson A., Schaechter M. Direct evidence for specific binding of the replicative origin of the Escherichia coli chromosome to the membrane. J Bacteriol. 1984 Apr;158(1):313–316. doi: 10.1128/jb.158.1.313-316.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Landoulsi A., Malki A., Kern R., Kohiyama M., Hughes P. The E. coli cell surface specifically prevents the initiation of DNA replication at oriC on hemimethylated DNA templates. Cell. 1990 Nov 30;63(5):1053–1060. doi: 10.1016/0092-8674(90)90508-c. [DOI] [PubMed] [Google Scholar]
- Nagai K., Hendrickson W., Balakrishnan R., Yamaki H., Boyd D., Schaechter M. Isolation of a replication origin complex from Escherichia coli. Proc Natl Acad Sci U S A. 1980 Jan;77(1):262–266. doi: 10.1073/pnas.77.1.262. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nicolaidis A. A., Holland I. B. Evidence for the specific association of the chromosomal origin with outer membrane fractions isolated from Escherichia coli. J Bacteriol. 1978 Jul;135(1):178–189. doi: 10.1128/jb.135.1.178-189.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Niki H., Jaffé A., Imamura R., Ogura T., Hiraga S. The new gene mukB codes for a 177 kd protein with coiled-coil domains involved in chromosome partitioning of E. coli. EMBO J. 1991 Jan;10(1):183–193. doi: 10.1002/j.1460-2075.1991.tb07935.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ogden G. B., Pratt M. J., Schaechter M. The replicative origin of the E. coli chromosome binds to cell membranes only when hemimethylated. Cell. 1988 Jul 1;54(1):127–135. doi: 10.1016/0092-8674(88)90186-9. [DOI] [PubMed] [Google Scholar]
- Wolf-Watz H., Masters M. Deoxyribonucleic acid and outer membrane: strains diploid for the oriC region show elevated levels of deoxyribonucleic acid-binding protein and evidence for specific binding of the oriC region to outer membrane. J Bacteriol. 1979 Oct;140(1):50–58. doi: 10.1128/jb.140.1.50-58.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]