Use of a transposon (Tndif) to obtain suppressing and nonsuppressing insertions of the dif resolvase site of Escherichia coli.
The dif locus is a RecA-independent recombination site, located in the terminus region of the chromosome of Escherichia coli. This site functions to reduce circular dimer chromosomes to monomers before cell division. Strains lacking this site exhibit the Dif phenotype, in which a fraction of the cells form extended filaments with abnormal nucleoids, and the SOS system is induced. We have used a transposon (Tndif), as well as linear transformation, to position dif in 19 locations around the chromosome. All of the suppressing insertions that we obtained were within 10 kb of the normal site, even in strains in which the normal symmetry, between the origin of replication and dif had been altered by 200 kb. We also observed that the nonsuppressing insertions in the terminus region became suppressing if a deletion occurred that extended from the ectopic site up to or past the normal location of dif. We propose that dif is normally located at the center of converging polarities in the terminus region and that deletions that restore suppression do so by placing ectopic sites once again at the center of this polarity. Similar results and conclusions are described in this issue.
Kuempel
P
P
Department of Molecular, Cellular, and Development Biology, University of Colorado, Boulder 80309, USA.
Høgaard
A
A
Nielsen
M
M
Nagappan
O
O
Tecklenburg
M
M
D002876:Chromosomes, Bacterial; D004254:DNA Nucleotidyltransferases; D004261:DNA Replication; D004251:DNA Transposable Elements / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics*; D008957:Models, Genetic; D016254:Mutagenesis, Insertional; D010957:Plasmids / Q000235:genetics; D011995:Recombination, Genetic / Q000235:genetics*; D017384:Sequence Deletion; D013489:Suppression, Genetic; D014169:Transformation, Bacterial; D019895:Transposases
D004251:DNA Transposable Elements; D004254:DNA Nucleotidyltransferases; D019895:Transposases