Studies on the role of dam methylation at the Escherichia coli chromosome replication origin (oriC).
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Biomedical subjects
Publications and source records attributed to P Forterre.
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Aphidicolin, a specific inhibitor of eucaryotic alpha DNA polymerase, inhibits the growth of halophilic arachaebacteria. In Halobacterium halobium, aphidicolin prevents cell division and DNA synthesis. These results suggest that arachaebacterial replicases are of the eucaryotic type.
We have examined the presence of methylated adenine at GATC sequences (Dam phenotype) in the DNA of 23 eubacteria and 13 archaebacteria by using isoshizomer restriction enzymes. We have found a completely Dam+ phenotype in bacteria of nine genera related to the families Enterobacteriaceae, Parvobacteriaceae, and Vibrionaceae, and in the five cyanobacteria tested. We have found a partial Dam+ phenotype in the two archaebacteria Halobacterium saccharovorum and Methanobacterium sp. strain Ivanov. All of the other archaebacteria (three genera) and eubacteria (nine genera) tested were Dam-. Phylogenetic analysis, based on the evolutionary tree of Fox et al. (Science 209:457-463, 1980), indicates that dam methylation in the Escherichia coli lineage appeared recently in bacterial evolution and is restricted to a small range of closely related bacteria.
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Toluene-treated Escherichia coli can synthesize DNA in the presence of precursors and ATP [Moses, R.E. & Richardson, C.C. (1970) Proc. Natl Acad. Sci. U.S.A. 67, 674--681]. The replacement of ATP by another NTP or dNTP leads to the premature arrest of the reaction. Residual synthesis in the presence of an NTP or dNTP other than ATP differs from the complete reaction in the presence of ATP because it is less sensitive to nalidixic acid and novobiocin and because its maximal activity can be obtained with lower concentrations of dNTP or shorter times of toluene treatment. However, like the complete reaction, residual synthesis occurs at the replication fork pre-existing in vivo at the time of toluenization, produces short and long pieces of DNA, is inhibited by arabinosyl-adenine triphosphate, azide or mitomycin C, and is dependent on the dnaE, DNAB and dnaG gene products. We conclude from these data that ATP is specifically required for a step in DNA replication which involves the activity of DNA gyrase, the target of nalidixic acid and novobiocin [Higgins, N.P., Peebles, C.L., Sugino, A. & Cozzarelli, N.R. (1978) Proc. Natl Acad. Sci. U.S.A. 75, 1773-1777]. In the absence of DNA gyrase activity, short DNA pieces are formed and sealed but only a limited amount of the chromosome can be replicated (residual synthesis). In the presence of DNA gyrase activity, DNA synthesis can occur on a longer portion of the chromosome (complete synthesis).
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Molecular phylogenetic studies have revealed a tripartite division of the living world into two procaryotic groups, Bacteria and Archaea, and one eucaryotic group, Eucarya. Which group is the most "primitive"? Which groups are sister? The answer to these questions would help to delineate the characters of the last common ancestor to all living beings, as a first step to reconstruct the earliest periods of biological evolution on Earth. The current "Procaryotic dogma" claims that procaryotes are primitive. Since the ancestor of Archaea was most probably a hyperthermophile, and since bacteria too might have originated from hyperthermophiles, the procaryotic dogma has been recently connected to the hot origin of life hypothesis. However, the notion that present-day hyperthermophiles are primitive has been challenged by recent findings, in these unique microorganisms, of very elaborate adaptative devices for life at high temperature. Accordingly, I discuss here alternative hypotheses that challenge the procaryotic dogma, such as the idea of a universal ancestor with molecular features in between those of eucaryotes and procaryotes, or the origin of procaryotes via thermophilic adaptation. Clearly, major evolutionary questions about early cellular evolution on Earth remain to be settled before we can speculate with confidence about which kinds of life might have appeared on other planets.