The tetracycline resistance determinants of RP1 and Tn<i>l721</i>: nucleotide sequence analysis

Waters, Simon; Rogowsky, Peter; Grinsted, J.; Altenbuchner, Josef; Schmitt, Rüdiger

doi:10.1093/nar/11.17.6089

Cited by 182 publications

(110 citation statements)

References 23 publications

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“…It is possible that as yet uncharacterized control elements lie elsewhere on RK2. Indeed, other weakly acting incompatibility loci have been located (Sakanyan et al, 1978;Meyer and Hinds, 1982 region as deduced from the DNA sequence Waters, 1982;Waters et al, 1983 The way in which copA and copB act is as yet unknown. Both of these elements express incompatibility suggesting that they are not simply cis-acting functions.…”

Section: Resultsmentioning

confidence: 99%

Analysis of copy number control elements in the region of the vegetative replication origin of the broad host range plasmid RK2.

Thomas¹,

Cross²,

Hussain³

et al. 1984

The EMBO Journal

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Broad host-range plasmid RK2 is able to replicate in a controlled manner in most Gram negative bacterial species. To analyze the elements of its control mechanism, we have measured the copy number in Escherichia coli of mini-RK2 replicons isogenic except for defined deletions in regions adjacent to the vegetative replication origin, ortVRK2, which have previously been implicated in copy number control because of their expression of plasmid incompatibility. The results indicate that while the previously defined 700-bp HaeII oriVRK2 fragment carries one copy control element (copA), a second (copB) lies at least partly outside this fragment towards the tetracycline resistance genes of RK2. Deletions affecting both these regions give a mini replicon with a copy number of 35-40 compared with 4-7 for parental RK2. Further incompatibility experiments indicate that targets for both incA (copA) and incB (copB) lie within the 700-bp HaeII oriVRK2 fragment.

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Section: Resultsmentioning

confidence: 99%

Analysis of copy number control elements in the region of the vegetative replication origin of the broad host range plasmid RK2.

Thomas¹,

Cross²,

Hussain³

et al. 1984

The EMBO Journal

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“…The line below the map indicates distance in base pairs and landmark restriction sites. The thin portion of the line shows the region whose sequence was known (53,61), while the thick portion indicates a 520 bp-region of the nucleotide sequence reported here. The TaqI site at position 801 corresponds to the TaqI site at position 1 in Fig.…”

Section: Methodsmentioning

confidence: 99%

Structure, expression, and regulation of the kilC operon of promiscuous IncP alpha plasmids

Larsen

Figurski

1994

J Bacteriol

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The kil-kor regulon was first identified on the broad-host-range IncPa plasmid RK2 by the presence of multiple kil loci (kiL4, kilB, kiC, and recently kiLE) that are lethal to Escherichia coli host cells in the absence of regulation by kor functions in various combinations. Whereas nine iterons of oriV. Iteron 10 is identical to the "orphan" iteron 1, and both have identical 6-bp flanking sequences that make them likely to be strong binding sites for the TrfA replication initiator protein. The locations and relative orientation of orphan iterons 10 and 1 raise the possibility that these iterons promote the formation of a DNA loop via protein-protein interactions by bound TrfA and lead us to propose that they demarcate the functional origin of replication. This analysis of the kilC region and our previous studies on the other kil loci of RK2 have revealed that the region between oriV and the korABF operon in wild-type IncPa plasmids is saturated by the kiC, kHlE, and kiL4 loci arranged in four kor-regulated operons encoding a total of 12 genes.The self-transmissible plasmids of incompatibility group P (IncPa and IncP,) are known for their conjugal and replicative promiscuity among diverse bacteria (10,40,42,55). Studies with the 60-kb IncPa plasmids RK2 (24) and RP4 (10) have revealed a unique regulatory network known as the kil-kor regulon (15,16,29). The eight operons of the kil-kor regulon are controlled by various combinations of transcriptional repressors encoded by korA, korB, and korC (15,29,55). The expression of several of the operons is further influenced by other regulatory functions encoded by korE, korFI, korFII, trbA, and kfrA (42,55,65). The functions of some of the proteins encoded by the kil-kor regulon are known (42). The trfA operon specifies the essential replication initiator function (TrfA), which binds to multiple copies of a 17-bp sequence (iteron) in the origin of replication (oriV), and a singlestranded DNA-binding protein (SSB). The korA operon includes the regulatory genes korA, korB, korFI and korFII, and

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“…Tet(B) and Tet(C), which are wellstudied Tet proteins in Gram-negative bacteria [16], are integral membrane proteins of about 43 kDa which consist of 12 membrane-spanning segments with a large central loop [17,18]. On the other hand, the molecular sizes of Tet(K) and Tet(L) are greater than those of Tet(B) and Tet(C).…”

Section: **Present Address: Exploratory Research Laboratories Tukubamentioning

confidence: 99%

Transmembrane glutamic acid residues play essential roles in the metal‐tetracycline/H⁺ antiporter of Staphylococcus aureus

Fujihira¹,

Kimura²,

Shiina³

et al. 1996

FEBS Letters

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; however, it has no transmembrane aspartyl residue. On the other hand, Tet(K) has three glutamyl residues, Glu-30, Glu-152 and Glu-397, in the putative transmembrane regions. In the present work, tet(K) gene was expressed in Escherichia coli and the transport activity was measured in everted membrane vesicles. When these glutamyl residues were replaced with Gin, the tetracycline transport activity was almost completely lost, indicating the important roles of these residues in Tet(K). In the case of Glu-397, even the charge-conserved mutation to Asp caused complete loss of the activity. On the other hand, the mutation of Glu-30 and Glu-152 to Asp resulted in significant retention of transport activity. These results are similar to those on the mutation of the three transmemhrane aspartyl residues in Tet(B), indicating that the transmembrane glutamyl residues in Tet(K) play roles similar to those of the transmemhrane aspartyl residues in Tet(B).

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The tetracycline resistance determinants of RP1 and Tnl721: nucleotide sequence analysis

Cited by 182 publications

References 23 publications

Analysis of copy number control elements in the region of the vegetative replication origin of the broad host range plasmid RK2.

Analysis of copy number control elements in the region of the vegetative replication origin of the broad host range plasmid RK2.

Structure, expression, and regulation of the kilC operon of promiscuous IncP alpha plasmids

Transmembrane glutamic acid residues play essential roles in the metal‐tetracycline/H⁺ antiporter of Staphylococcus aureus

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The tetracycline resistance determinants of RP1 and Tnl721: nucleotide sequence analysis

Cited by 182 publications

References 23 publications

Analysis of copy number control elements in the region of the vegetative replication origin of the broad host range plasmid RK2.

Analysis of copy number control elements in the region of the vegetative replication origin of the broad host range plasmid RK2.

Structure, expression, and regulation of the kilC operon of promiscuous IncP alpha plasmids

Transmembrane glutamic acid residues play essential roles in the metal‐tetracycline/H+ antiporter of Staphylococcus aureus

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Transmembrane glutamic acid residues play essential roles in the metal‐tetracycline/H⁺ antiporter of Staphylococcus aureus