Copy mutants of plasmid R1: effects of base pair substitutions in the copA gene on the replication control system

M Givskov, Søren Molin

26 Citations (Scopus)

Abstract

Five different copA copy mutants of plasmid R1 have been identified by nucleotide sequencing. Independent measurements of the activities of the mutant inhibitor RNA and of the mutant target properties were carried out using several different methods. Correlation of these measurements with the location of th nucleotide substitutions resulted in the following conclusions: (1) The copy number of plasmid R1 is controlled primarily by interaction between the CopA RNA molecule and its target, the RepA mRNA. (2) The binding of th inhibitor to its target is based on nucleotide interactions within two complementary sequences of ten nucleotides and dependent on the secondary structure of the active site. (3) The secondary structure of both the CopA target and the CopA RNA is a stem-loop structure. Mutations in the loop region interfere with binding affinity between inhibitor and target, whereas mutations in the upper stem mainly interfere with secondary structure. Mutations in the latter region create temperature-dependent copy number phenotypes.
Original languageEnglish
JournalMolecular and General Genetics MGG
Volume194
Issue number1-2
Pages (from-to)286-92
Number of pages7
ISSN0026-8925
Publication statusPublished - 1984
Externally publishedYes

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