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Biomedical subjects

S Z Mindlin

Publications and source records attributed to S Z Mindlin.

At least 19 recordsLinked to original sources

[Origin, evolution, and migration of drug resistance genes].

Current views on the mechanisms responsible for the emergence of multiple drug resistance in clinical bacterial isolates are considered. Hypotheses on the origin of resistance genes derived from determinants of actinomycetes, antibiotic producers, and chromosomal genes of bacteria involved in cellular metabolism are reviewed. The mechanisms underlying the diffusion of resistance determinants by means of bacterial mobile elements (plasmids, transposons, and integrons) are discussed. Examples of the horizontal transfer of resistance determinants between Gram-positive and Gram-negative bacteria are presented.

Amino Acid Sequence↗

[Distribution of transposons Tn5044 and Tn5070 with unusual mer operons in environmental bacterial populations].

The distribution of unusual mercury resistance transposons, Tn5044 and Tn5070, was examined. A characteristic feature of Tn5044 is temperature sensitivity of its mercury operon and the presence in the mer operon of the gene homologous to RNA polymerase a subunit. Structural organization of mercury operon Tn5070, containing minimum gene set (merRTPA), differs from mer operons of both Gram-negative and Gram-positive bacteria. None of more than two thousand environmental bacterial strains displaying mercury resistance and isolated from the samples selected from different geographical regions hybridized to Tn5040- and Tn5070-specific probes. A concept on the existence of cosmopolite, endemic, and rare transposons in environmental bacterial populations was formulated.

DNA Transposable Elements↗

[Mercury-resistant bacteria from permafrost sediments and prospects for their use in comparative studies of mercury resistance determinants].

Mercury-resistant bacteria were isolated from permafrost sediments of Kolyma lowland and Canada existing over five thousand to two million years. Their content was shown to vary within the range 0.001-2.9% and to depend on the amount of mercury in sampling sites (coefficient of correlation 0.75). A collection of mercury-resistant bacterial strains was created. In this collection, various representatives of both Gram-positive bacteria (Bacillus, Exiguobacterium, Micrococcus, Arthrobacter) and Gram-negative bacteria (Pseudomonas, Acinetobacter, Plesiomonas, Myxobacteriales) were identified. Most resistant bacteria were found to contain determinants homologous to mer-operons of contemporary bacteria. The isolated strains of paleobacteria are proposed to be used for a comparative structural study of contemporary and ancient plasmids and transposons carrying mercury resistance determinants.

Bacteria↗

[Molecular genetic analysis of the Tn5041 transposition system].

A study was made of the transposition of the mercury resistance transposon Tn5041 which, together with the closely related toluene degradation transposon Tn4651, forms a separate group in the Tn3 family. Transposition of Tn5041 was host-dependent: the element transposed in its original host Pseudomonas sp. KHP41 but not in P. aeruginosa PAO-R and Escherichia coli K12. Transposition of Tn5041 in these strains proved to be complemented by the transposase gene (tnpA) of Tn4651. The gene region determining the host dependence of Tn5041 transposition was localized with the use of a series of hybrid (Tn5041 x Tn4651) tnpA genes. Its location in the 5'-terminal one-third of the transposase gene is consistent with the data that this region is involved in the formation of the transposition complex in transposons of the Tn3 family. As in other transposons of this family, transposition of Tn5041 occurred via cointegrate formation, suggesting its replicative mechanism. However, neither of the putative resolution proteins encoded by Tn5041 resolved the cointegrates formed during transposition or an artificial cointegrate in E. coli K12. Similar data were obtained with the mercury resistance transposons isolated from environmental Pseudomonas strains and closely related to Tn5041 (Tn5041 subgroup).

Base Sequence↗

Horizontal spread of mer operons among gram-positive bacteria in natural environments.

Horizontal dissemination of the genes responsible for resistance to toxic pollutants may play a key role in the adaptation of bacterial populations to environmental contaminants. However, the frequency and extent of gene dissemination in natural environments is not known. A natural horizontal spread of two distinct mercury resistance (mer) operon variants, which occurred amongst diverse Bacillus and related species over wide geographical areas, is reported. One mer variant encodes a mercuric reductase with a single N-terminal domain, whilst the other encodes a reductase with a duplicated N-terminal domain. The strains containing the former mer operon types are sensitive to organomercurials, and are most common in the terrestrial mercury-resistant Bacillus populations studied in this work. The strains containing the latter operon types are resistant to organomercurials, and dominate in a Minamata Bay mercury-resistant Bacillus population, previously described in the literature. At least three distinct transposons (related to a class II vancomycin-resistance transposon, Tn1546, from a clinical Enterococcus strain) and conjugative plasmids are implicated as mediators of the spread of these mer operons.

Bacillus↗

Tn5041: a chimeric mercury resistance transposon closely related to the toluene degradative transposon Tn4651.

This paper reports the discovery and characterization of Tn5041, a novel-type transposon vehicle for dissemination of mercury resistance in natural bacterial populations. Tn5041 (14876 bp), identified in a Pseudomonas strain from a mercury mine, is a Tn3 family mercury resistance transposon far outside the Tn21 subgroup. As in other Tn3 family transposons, Tn5041 duplicates 5 bp of the target sequence following insertion. Tn5041 apparently acquired its mer operon as a single-ended relic of a transposon belonging to the classical mercury resistance transposons of the Tn21 subgroup. The putative transposase and the 47 bp terminal inverted repeats of Tn5041 are closely related to those of the toluene degradative transposon Tn4651 and fall into a distinct subgroup on the fringe of the Tn3 family. The amino acid sequence of the putative resolvase of Tn5041 resembles site-specific recombinases of the integrase family. Besides the mer operon and putative transposition genes, Tn5041 contains a 4 kb region that accommodates a number of apparently defective genes and mobile elements.

Amino Acid Sequence↗

[Incorporation of Tn5053 and Tn402 into various plasmids].

Transposons Tn5053 and Tn402 that belong to the novel family of Tn elements are characterized by high selectivity when choosing a target. These transposons integrated with a high frequency into only two of seven large plasmids of various incompatibility groups: RP1 and R446b. The res region of the RP1 plasmid par locus and the res region of the transposon Tn701, included into R446b plasmid, served as targets for both transposons. When Tn701 or par locus integrated into plasmids previously unsuitable for Tn5053 and Tn402 transposition, these plasmids became good targets for both transposons. On the contrary, when the res region of RP1 was damaged impaired, this good target became unsuitable. The insertion sites of Tn5053 and Tn402 were concentrated in the res region of Tn1721 and RP1, but, in some cases, they were at a distance of 100-2000 bp from it.

Base Sequence↗

Four genes, two ends, and a res region are involved in transposition of Tn5053: a paradigm for a novel family of transposons carrying either a mer operon or an integron.

The complete nucleotide sequence of an 8447 bp-long mercury-resistance transposon (Tn5053) has been determined. Tn5053 is composed of two modules: (i) the mercury-resistance module and (ii) the transposition module. The mercury-resistance module carries a mer operon, merRTPFAD, and appears to be a single-ended relic of a transposon closely related to the classical mercury-resistance transposons Tn21 and Tn501. The transposition module of Tn5053 is bounded by 25 bp terminal inverted repeats and contains four genes involved in transposition, i.e. tniA, tniB, tniQ, and tniR. Transposition of Tn5053 occurs via cointegrate formation mediated by the products of the tniABQ genes, followed by site-specific cointegrate resolution. This is catalysed by the product of the tniR gene at the res region, which is located upstream of tniR. The same pathway of transposition is used by Tn402 (Tn5090) which carries the integron of R751. Transposition genes of Tn5053 and Tn402 are interchangeable. Sequence analysis suggests that Tn5053 and Tn402 are representatives of a new family of transposable elements, which fall into a recently recognized super-family of transposons including retroviruses, insertion sequences of the IS3 family, and transposons Tn552 and Tn7. We suggest that the tni genes were involved in the dissemination of integrons.

Amino Acid Sequence↗

Tn5053, a mercury resistance transposon with integron's ends.

We describe a novel type of mercury resistance transposon, Tn5053, which was found in the chromosome of a mercury-resistant Xanthomonas strain isolated from a mercury mine. An 8400 base-pair Tn5053 is bracketed by 25 base-pair inverted repeats that have no sequence homology with inverted repeats of classical mercury resistance transposons Tn501 and Tn21. Instead they show high homology with inverted repeats bracketing the antibiotic resistance segment of Tn21 (integron In2). A 38 base-pair element, which is highly homologous to the inverted repeats of classical mercury resistance transposons has been found within Tn5053 near one of its ends. This internal inverted repeat is fused to the mer operon of Tn5053 in exactly the same way as in the Tn501 mercury resistance transposon. This finding suggests that the mer operon was integrated into the Tn5053 transposition module not through integron-specific pathway but rather via insertion of a classical mercury resistance transposon.

Base Sequence↗

Molecular characterization of an aberrant mercury resistance transposable element from an environmental Acinetobacter strain.

We present the complete nucleotide sequence of a mer operon located on a 60-kb conjugative plasmid pKLH2 from an environmental bacterium, Acinetobacter calcoaceticus, isolated from a mercury mine. The pKLH2 mer operon has essentially the same gene organization as that of Tn21 and Tn501 from clinical bacteria. The pKLH2 mer operon nucleotide sequence shows 85.5% identity with the Tn501 and 80.9% identity with the Tn21 sequences. Vestigial sequences have been found at the ends of the pKLH2 mer operon, indicating that the pKLH2 mer operon was once a part of a Tn21-like transposon, which had committed suicide by an aberrant resolution event.

Acinetobacter↗

Mercuric reductase in environmental gram-positive bacteria sensitive to mercury.

According to existing data, mercury resistance operons (mer operons) are in general thought to be rare in bacteria, other than those from mercury-contaminated sites. We have found that a high proportion of strains in environmental isolates of Gram-positive bacteria express mercuric reductase (MerA protein): the majority of these strains are apparently sensitive to mercury. The expression of MerA was also inducible in all cases. These results imply the presence of phenotypically cryptic mer resistance operons, with both the merA (mercuric reductase) and merR (regulatory) genes still present, but the possible absence of the transport function required to complete the resistance mechanism. This indicates that mer operons or parts thereof are more widely spread in nature than is suggested by the frequency of mercury-resistant bacteria.

Drug Resistance, Microbial↗

Occurrence of two structural types of mercury reductases among gram-positive bacteria.

Structural variants of mercury reductase containing the N-terminal domain, which is easily cleaved by trypsin, have been found in Gram-positive bacteria with a low genomic G + C content (Bacillus, Staphylococcus and, possibly, some other genera). Mercury reductases without the N-terminal domain and relatively resistant to limited proteolysis are typical for Gram-positive bacteria with a high genomic G + C content (Arthrobacter, Citreobacterium, Micrococcus, Mycobacterium, Rhodococcus). Both types of mercury reductase genes may be located on plasmids.

Base Composition↗

[Spontaneous transformation in mixed cultures of various types of Acinetobacter and during joint growth of Acinetobacter calcoaceticus with Escherichia coli and Pseudomonas aeruginosa].

The transfer of chromosomal and plasmid genes was studied via spontaneous transformation is mixed cultures of Acinetobacter spp. It turned out that any Acinetobacter strain, irrespective of its species specificity, serves as chromosomal DNA donor in case the mixed culture contains competent cells of the recipient strain. No transfer took place when non-related bacteria were used as donors. We also studied the transfer into Ac. calcoaceticus competent strain cells of small non-conjugative plasmids having broad host range (RSF1010, pAK1). In these cases, DNA donors could be not only acinetobacters of other species, but bacteria belonging to other systematic groups (families)--E. coli and P. aeruginosa. The transfer of plasmids from cells of unrelated bacteria took place with a frequency of about 10(-5)-10(-6). The possible role of spontaneous transformation in horizontal gene transfer is discussed.

Acinetobacter↗

Two structural types of mercury reductases and possible ways of their evolution.

Structural differences have been revealed among mercury reductases of immunologically unrelated types from Gram-positive bacteria: enzymes of one immunological type have a molecular mass of 62-69 kDa and seem to contain an N-terminal extension of 5-15 kDa, which is easily cleaved by trypsin and chymotrypsin; enzymes of the other immunological type have a molecular mass of 52-57 kDa and are resistant to proteolysis. The existence of at least two different lines in the evolution of mercury reductases is surmised.

Arthrobacter↗

The diversity of mercury reductases among mercury-resistant bacteria.

Two immunologically non-cross-reactive types of mercury reductases were found among Gram-negative and two among Gram-positive mercury-resistant environmental bacteria. Mercury reductases were further discriminated by 'spur' formation immunodiffusion tests. Immunologically indistinguishable mercury reductases were found among strains belonging to phylogenetically distant genera. This suggests a horizontal transfer of mercury resistance genes between these strains.

Bacteria↗

[Rpo pathway as a possible recombination mechanism of the interaction between DNA of transducing phages and the Escherichia coli genome].

The conditions affecting recombination of DNAs of transducing lambdoid rifd phages with the chromosome of Escherichia coli K-12 recA in the region of homology were studied. In support to the previously obtained data, the Int system was shown to take no part in the process. The homologous character of recombination interactions and their dependence on efficiency of transcription were demonstrated. It is therefore suggested that recombination takes the Rpo pathway. Similar peculiarities were revealed in the processes of interaction between DNAs of rifd phages as well as of lambdoid phages carrying trp genes and the host genome. A hypothesis is put forward that the Rpo pathway operates, depending on the density of DNA supercoiling.

Bacteriophage lambda↗

[Study of the horizontal transfer of mercury resistance genes in natural populations of bacteria using antibodies to mercury reductases].

Mercury resistant soil and intestinal bacteria were isolated from different mercury deposit areas of the USSR. Mercury reductases from all gram negative bacteria studied (Pseudomonas, Acinetobacter and Enterobacterial species) with a single exception (Flavobacterium sp.) were immunologically cross reactive. Two immunological types of mercury reductases were found among gram positive bacteria (Bacillus, Staphylococcus and Coryneform species). Further subdivisions were done by "spur" formation tests. Despite considerable diversity of mercury reductases revealed in this study, we found several strains which belonged to distant genera but contained immunologically indistinguishable enzymes. This suggested that the horizontal spread of the corresponding genes occurred in these genera in relatively recent time.

Antibodies, Monoclonal↗

Integration of bacteriophages lambda and phi 80 in wild-type Escherichia coli at secondary attachment sites. II. Genetic structure and mechanism of polylysogen formation for lambda, phi 80 and the lambda att80 hybrid.

The frequency of occurrence and the genetic structure of polylysogens were studied for phages lambda, phi 80 and lambda att80. In the case of lambda, frequency of polylysogenization is high (0.20 to 0.41) with a tandem integration of prophages at the primary att site (att lambda). With phi 80 and lambda att80, this frequency is about 10 times lower, and usually one prophage becomes integrated at the primary att site (att80-I) while another (sometimes two others) integrates at one of the secondary sites. At least four secondary att80 sites have been found in wild-type Escherichia coli, two of which (near the his and tolC loci) are preferred. The frequency of secondary integration of phi 80 and lambda att80 does not differ significantly in the wild-type host and in that deleted for the primary att site (0.041 and 0.045, respectively, among surviving cells at an MOI of 10). Homoimmune superinfection has revealed a constitutive cI-independent expression of the phi 80 int gene in the prophage state. The only phi 80 tandem detected proved to be unstable. With the phi 80int- mutant, we observed stabilization of phi 80 tandems; as a consequence, their frequency of occurrence during coinfection with phi 80int+ was up to the lambda level and no nontandem insertions were found. A model is proposed for the phi 80 and lambda att80 nontandem integration.

Bacteriophage lambda↗