Complete nucleotide sequence and gene organization of the broad-host-range plasmid RSF1010

Scholz, Peter; Häring, Volker; Wittmann‐Liebold, Brigitte; Ashman, Keith; Bagdasarian, Michael; Scherzinger, Eberhard

doi:10.1016/0378-1119(89)90273-4

Cited by 361 publications

(317 citation statements)

References 21 publications

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“…The first is attributed to a chromosomal mutation that results in the alteration of the target sites for binding of streptomycin on bacterial ribosomes (Chang and Flaks 1972). The second is mediated by resistant (R) plasmids (Scholz et al 1989) that carry determinants homologous to strA-strB genes of plasmid RSF1010. These genes encode the streptomycin modifying enzymes aminoglycoside-3-phosphotransferase and aminoglycoside-6-phosphotransferase.…”

Section: Introductionmentioning

confidence: 99%

Characterization of plasmids that encode streptomycin‐resistance in bacterial epiphytes of apple

Huang

Burr

1999

Journal of Applied Microbiology

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T .-C. H UA NG A ND T. J . B UR R . 1999. Streptomycin resistance in strains of Pseudomonas syringae pv. papulans, Pantoea agglomerans and a yellow-pigmented, nonfluorescent Pseudomonas sp. (Py), isolated from apple orchards in New York and Washington states, is predominantly associated with strA-strB genes carried on conjugal plasmids (R plasmids). None of 128 resistant Erwinia amylovora strains from the eastern and western USA hybridized with a strA-strB probe, SMP3. Resistant Py strains transfered R plasmids to Ps. syringae pv. papulans and to Py in vitro at frequencies of 10 −1 -10 −2 per recipient cell whereas Ps. syringae pv. papulans transferred its plasmids at frequencies of 10 −2 to below detectable levels. Transfer of R plasmids to P. agglomerans was not detected and resistant P. agglomerans did not transfer their R plasmids to any recipients. R plasmids were found to be highly diverse as measured by DNA fingerprint analysis. Transfer-deficient transposon mutants of R plasmid pCPP519 were generated, and 3·9 kb EcoRI and 3·0 kb SmaI fragments that hybridized with a Tn5 probe were cloned and sequenced. The deduced amino acid sequences of the 3·9 kb fragment were similar to proteins involved in replication, nicking at oriT, and piliation in other bacteria.

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Section: Introductionmentioning

confidence: 99%

Characterization of plasmids that encode streptomycin‐resistance in bacterial epiphytes of apple

Huang

Burr

1999

Journal of Applied Microbiology

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“…RepBЈ recognizes ssiA and ssiB independently of each other (6, 7) and synthesizes primers (10) that are elongated by DNA polymerase III. Replication of RSF1010 depends on the ssi sequences that occur only once on each plasmid strand, suggesting that RSF1010 (11,12) is replicated in the leading-strand mode in opposite directions along single-stranded template DNA (10,13,14) (Fig. S1 A).…”

mentioning

confidence: 99%

Structure and function of primase RepB′ encoded by broad-host-range plasmid RSF1010 that replicates exclusively in leading-strand mode

Geibel

Banchenko

Engel

et al. 2009

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

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For the initiation of DNA replication, dsDNA is unwound by helicases. Primases then recognize specific sequences on the template DNA strands and synthesize complementary oligonucleotide primers that are elongated by DNA polymerases in leading-and lagging-strand mode. The bacterial plasmid RSF1010 provides a model for the initiation of DNA replication, because it encodes the smallest known primase RepB (35.9 kDa), features only 1 single-stranded primase initiation site on each strand (ssiA and ssiB, each 40 nt long with 5 -and 3 -terminal 6 and 13 single-stranded nucleotides, respectively, and nucleotides 7-27 forming a hairpin), and is replicated exclusively in leading strand mode. We present the crystal structure of full-length dumbbell-shaped RepB consisting of an N-terminal catalytic domain separated by a long ␣-helix and tether from the C-terminal helixbundle domain and the structure of the catalytic domain in a specific complex with the 6 5 -terminal single-stranded nucleotides and the C7-G27 base pair of ssiA, its single-stranded 3 -terminus being deleted. The catalytic domains of RepB and the archaeal/eukaryotic family of Pri-type primases share a common fold with conserved catalytic amino acids, but RepB lacks the zinc-binding motif typical of the Pri-type primases. According to complementation studies the catalytic domain shows primase activity only in the presence of the helix-bundle domain. Primases that are highly homologous to RepB are encoded by broad-host-range IncQ and IncQ-like plasmids that share primase initiation sites ssiA and ssiB and high sequence identity with RSF1010.Pri-type ͉ primase DNA complex ͉ single-strand initiator DNA ͉ crystal structure ͉ oriV A t the initiation of DNA replication, the double helix is unwound by helicases, and then primases synthesize complementary oligonucleotide primers on the single-stranded template DNA. These primers are extended by DNA polymerase, continuously at the leading strand and discontinuously at the oppositely-oriented lagging strand, where Okazaki fragments are synthesized. Primase activity depends on specific ssDNA priming sequences that are periodically required for the initiation of the Okazaki fragments on the lagging strand (that are connected by ligase) but only 1 primer is used for the leading strand (1).The broad-host-range IncQ plasmid RSF1010 is maintained in Ͼ30 Gram-negative and 2 Gram-positive bacteria (2, 3). The replication of IncQ and IncQ-like plasmids and their conjugative transfer was studied in Escherichia coli and in vitro. RSF1010 encodes helicase RepA, primase RepBЈ, and replication initiator protein RepC (4, 5) (Fig. S1 A). Additionally, the replication of RSF1010 requires bacterial host replication proteins such as DNA polymerase III and single-stranded binding protein.Primase RepBЈ is activated on 2 specific, 40-nt-long singlestranded segments termed initiators A and B (ssiA and ssiB) with highly similar nucleotide sequences (6, 7). They are arranged on opposite DNA strands as a palindrome in the origin of vegetative re...

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“…IncQ-type plasmid replicons contain three replication genes, repA, repB and repC, and an oriV region (Scholz et al, 1989). In the case of the best-studied IncQ-1a plasmids, RSF1010 and R1162, the oriV region contains three and one half 20 bp iterons with 2 bp nucleotide spacers.…”

Section: Introductionmentioning

confidence: 99%

Evolution of compatible replicons of the related IncQ-like plasmids, pTC-F14 and pTF-FC2

Gardner

Rawlings

2004

Microbiology

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Two closely related but compatible plasmids of the IncQ-2a and IncQ-2b groups, pTF-FC2 and pTC-F14, were discovered in two acidiphilic chemolithotrophic bacteria. Cross-complementation and cross-regulation experiments by the replication proteins were carried out to discover what changes were necessary when the plasmids evolved to produce two incompatibility groups. The requirement of a pTC-F14 oriV for a RepC DNA-binding protein was plasmid specific, whereas the requirement for the RepA helicase and RepB primase was less specific and could be complemented by the IncQ-2a plasmid pTC-FC2, and the IncQ-1b plasmid pIE1108. None of the IncQ-1a plasmid replication proteins could complement the pTC-F14 oriV, and pTC-F14 and RSF1010 were incompatible. This incompatibility was associated with the RepC replication protein and was not due to iteron incompatibility. Replication of pTC-F14 took place from a 5?7 kb transcript that originated upstream of the mobB gene located within the region required for mobilization. A pTC-F14 mobB-lacZ fusion was regulated by the pTC-F14 repB gene product and was plasmid specific, as it was not regulated by the RepB proteins of pTF-FC2 or the IncQ-1a and IncQ-1b plasmids. Plasmid pTC-F14 appears to have evolved independently functioning iterons and a plasmid-specific RepC-binding protein; it also has a major replication transcript that is independently regulated from that of pTF-FC2. However, the RepA and RepB proteins have the ability to function with either replicon.

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Complete nucleotide sequence and gene organization of the broad-host-range plasmid RSF1010

Cited by 361 publications

References 21 publications

Characterization of plasmids that encode streptomycin‐resistance in bacterial epiphytes of apple

Characterization of plasmids that encode streptomycin‐resistance in bacterial epiphytes of apple

Structure and function of primase RepB′ encoded by broad-host-range plasmid RSF1010 that replicates exclusively in leading-strand mode

Evolution of compatible replicons of the related IncQ-like plasmids, pTC-F14 and pTF-FC2

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