Mutations in the DnaA binding sites of the replication origin of Escherichia coli. |
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Authors: | Andreas Holz Christoph Schaefer Hendrik Gille Wolf-Rainer Jueterbock Walter Messer |
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Affiliation: | (1) Max-Planck-Institut für Molekulare Genetik, Ihnestrasse 73, W-1000 Berlin 33, Germany;(2) Present address: Institute of Brain Research, August Forel Strasse 1, CH-8029 Zürich, Schweiz;(3) Present address: MPI f. Immunbiologie, Spemann Laboratories, Postfach 1169, W-7800 Freiburg-Zähringen, Germany |
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Abstract: | Summary Mutations (base changes) were introduced into the four DnaA binding sites (DnaA boxes) of theEscherichia coli replication origin,oriC. Mutations in a single DnaA box did not impair the ability of these origins to replicate in vivo and in vitro. A combination of mutations in two DnaA boxes, R1 and R4, resulted in slower growth of theoriC plasmid-bearing host cells. DnaA protein interaction with mutant and wild-type DnaA boxes was analyzed by DNase I footprinting. Binding of DnaA protein to a mutated DnaA box R1 was not affected by a mutation in DnaA box R4 and vice versa. Mutations in DnaA boxes R1 and R4 did not modify the ability of the DnaA protein to bind to other DnaA boxes inoriC. |
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Keywords: | Initiation of replication Initiation complex oriC |
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