Characterization and DNA sequence of the mobilization region of pLV22a from Bacteroides fragilis.
A 4.2-kb plasmid (pLV22a) native to Bacteroides fragilis LV22 became fused to a transfer-deficient Bacteroides spp.-Escherichia coli shuttle vector by an inverse transposition event, resulting in a transferrable phenotype. The transfer phenotype was attributable to pLV22a, which was also capable of mobilization within E. coli when coresident with the IncP beta R751 plasmid. Transposon mutagenesis with Tn1000 localized the mobilization region to a 1.5-kb DNA segment in pLV22a. The mobilization region has been sequenced, and five open reading frames have been identified. Mutants carrying disruptions in any of the three genes designated mbpA, mbpB, and mbpC and coding for deduced products of 11.3, 30.4, and 17.1 kDa, respectively, cannot be mobilized when coresident with R751. Mutations in all three genes can be complemented in the presence of the respective wild-type genes, indicating that the products of mbpA, mbpB, and mbpC have roles in the mobilization process and function in trans. The deduced 30.4-kDa MbpB protein contains a 14-amino-acid conserved motif that is also found in the DNA relaxases of a variety of conjugal and mobilizable plasmids and the conjugative transposon Tn4399. Deletion analysis and complementation experiments have localized a cis-acting region of pLV22a within mbpA.
Novicki
T J
TJ
Department of Medicine, VA Hospital, Hines, Illinois 60141, USA.
Hecht
D W
DW
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics; D001441:Bacteroides fragilis / Q000235:genetics*; D001483:Base Sequence; D003227:Conjugation, Genetic; D004264:DNA Topoisomerases, Type I / Q000235:genetics; D004251:DNA Transposable Elements; D015964:Gene Expression Regulation, Bacterial; D005816:Genetic Complementation Test; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D016366:Open Reading Frames; D010957:Plasmids / Q000235:genetics*; D017386:Sequence Homology, Amino Acid; D014169:Transformation, Bacterial
D001426:Bacterial Proteins; D004251:DNA Transposable Elements; C095095:MbpA protein, Bacteroides fragilis; C095096:MbpB protein, Bacteroides fragilis; C095097:MbpC protein, Bacteroides fragilis; D004264:DNA Topoisomerases, Type I