Deletion derivatives of pAgK84 and their use in the analysis of Agrobacterium plasmid functions

Farrand, Stephen K.; Hong, Seung‐Beom; O’Morchoe, S. B.; Slota, John E.

doi:10.1016/0147-619x(92)90052-c

Cited by 9 publications

(7 citation statements)

References 35 publications

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“…Moreover, because orf11-orf12-stb may form a transcriptional unit, it is likely that the transposon insertion abolished expression of this gene set. This hypothesis is consistent with the fact that Tn5 insertions in the orf11-orf12-stb region have no effect on plasmid stability (16). We suspect that pAgBo542 is not ancestral but arose from an event in which this 7-kb region had been excised.…”

Section: Resultssupporting

confidence: 74%

“…The stb product contains a ParB domain and is related to a putative stabilization protein from Rhodopseudomonas palustris. The location of these genes is consistent with deletion analyses that yielded a minimum stable replicon containing this segment of the plasmid (16). Two small ORFs, orf11 and orf12, are located immediately upstream of stb, and could be part of the same transcriptional unit.…”

Section: Resultssupporting

confidence: 54%

“…The 42-bp intergenic region between orf12 and stb contains neither a good rho-independent terminator nor identifiable promoter sequences. However, transposon insertions mapping within the orf11-stb region have no effect on plasmid stability (16), suggesting that, if this gene set is expressed, neither stb nor orfs 11 and 12 are essential for replication or efficient partitioning (see below).…”

Section: Resultsmentioning

confidence: 99%

“…Genes for synthesis of and immunity to the antibiotic map to five complementation groups covering a contiguous 15-kb region of the plasmid (13)(14)(15)). An additional 8 kb is required for replication and transfer (13,16). Here, we present the entire nucleotide sequence of pAgK84.…”

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confidence: 99%

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Bases of biocontrol: Sequence predicts synthesis and mode of action of agrocin 84, the Trojan Horse antibiotic that controls crown gall

Kim

Park

Kim

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

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Agrobacterium radiobacter K84, used worldwide to biocontrol crown gall disease caused by Agrobacterium tumefaciens, produces an antiagrobacterial compound called agrocin 84. We report the nucleotide sequence of pAgK84, a 44.42-kb plasmid coding for production of this disubstituted adenine nucleotide antibiotic. pAgK84 encodes 36 ORFs, 17 of which (agn) code for synthesis of or immunity to agrocin 84. Two genes, agnB2 and agnA, encode aminoacyl tRNA synthetase homologues. We have shown that the toxic moiety of agrocin 84 inhibits cellular leucyl-tRNA synthetases and AgnB2, which confers immunity to the antibiotic, is a resistant form of this enzyme. AgnA, a truncated homologue of asparaginyl tRNA synthetase could catalyze the phosphoramidate bond between a precursor of the methyl pentanamide side group and the nucleotide. We propose previously undescribed chemistry, catalyzed by AgnB1, to generate the precursor necessary for this phosphoramidate linkage. AgnC7 is related to ribonucleotide reductases and could generate the 3 -deoxyarabinose moiety of the nucleoside. Bioinformatics suggest that agnC3, agnC4, and agnC6 contribute to maturation of the methyl pentanamide, whereas agnC2 may produce the glucofuranose side group bound to the adenine ring. AgnG is related to bacterial exporters. An agnG mutant accumulated agrocin 84 intracellularly but did not export the antibiotic. pAgK84 is transmissible and encodes genes for conjugative DNA processing but lacks a type IV secretion system, suggesting that pAgK84 transfers by mobilization. By sequence analysis, the deletion engineered into pAgK1026 removed the oriT and essential tra genes, confirming the enhanced environmental safety of this modified form of pAgK84.Agrobacterium radiobacter ͉ biological control ͉ pAgK84

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Section: Resultssupporting

confidence: 74%

Section: Resultssupporting

confidence: 54%

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

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Bases of biocontrol: Sequence predicts synthesis and mode of action of agrocin 84, the Trojan Horse antibiotic that controls crown gall

Kim

Park

Kim

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

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“…On the other hand, matings with recipients outside of the family Rhizobiaceae do not yield stable transconjugants. However, genetic analysis indicates that no more than 3 kb of this plasmid is responsible for conjugation (Farrand, et al, 1985;Farrand, et al, 1992). Some years ago Sprinzel and Geider (1988) showed, using a zygotic induction assay, that pTiB6 is transmissible to E. coli recipients.…”

Section: Ye Conmgal Host Rangementioning

confidence: 99%