Genetic Structure and DNA Sequences at Junctions Involved in the Rearrangements of Bacillus Subtilis Strains Carrying the Trpe26 Mutation

Studies on the region upstream to ribosomal operon rrnD of Bacillus subtilis led to the characterization of two of the four chromosomal junctions involved in the rearrangements (a translocation and an inversion) of the strains carrying the trpE26 mutation. Genetic analysis, by integrative mapping, showed linkage of rrnD to cysB and hisA (both on segment A) in the trpE26-type strains. Physical analysis showed that the region upstream to rrnD is now linked to the trpE-ilvA chromosome segment as demonstrated by analyzing restriction site-polymorphism between 168 and trpE26-type strains. Similar experiments confirmed the previous genetic data on linkage in these areas in strains carrying novel rearrangements derived from the trpE26-type strains: stable merodiploids and inversions. The nucleotide sequence of the area 5' to rrnD in both types of strains (168 and trpE26), the region downstream of the citG gene and the region carrying the trpE26 mutation (made available to us by D. Henner) provided evidence for the molecular basis of the differences in structure, allowed the identification of the break points and revealed the presence of a polypurine region upstream to rrnD as seen in other systems in B. subtilis. No extensive homology was found between pairs of junctions so far sequenced. The models proposed by C. Anagnostopoulos for the role of DNA sequences of intrachromosomal homology involved in the transfer of the trpE26 mutation and the formation of novel arrangements require therefore reevaluation.