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S M Mirkin

Publications and source records attributed to S M Mirkin.

At least 37 records · Page 2Linked to original sources
PubMed1988

Chemical probing of homopurine-homopyrimidine mirror repeats in supercoiled DNA.

We have recently shown that under superhelical stress and/or acid pH the homopurine-homopyrimidine tracts conforming to the mirror symmetry (H palindromes) form a novel DNA structure, the H form. According to our model, the H form includes (1) a triplex formed by half of the purine strand and by the homopyrimidine hairpin and (2) the unstructured other half of the purine strand. We used four specially designed sequences to demonstrate that, in accordance with our model, the mirror symmetry is essential for facile formation of the H form detected by two-dimensional gel electrophoresis. Here we report that, under conditions favouring the H-form extrusion, guanines of the 3' half of the purine strand are protected against alkylation by dimethylsulphate, whereas adenines of the 5' half of the purine strand react with diethyl pyrocarbonate. These data indicate that the 3' half of the homopurine strand is within the triplex whereas the 5' half is unstructured, in full agreement with our model.

DNA, Superhelical↗
PubMed1988

A stable complex between homopyrimidine oligomers and the homologous regions of duplex DNAs.

When plasmid DNA duplexes carrying the regular homopurine-homopyrimidine inserts (dGA)n, (dTC)n and (dG)n, (dC)n are preincubated with homologous labeled oligo(dPy) ((dTC)n and (dC)n respectively) at acid pH, the label co-electrophoreses with the duplex DNA. Thus, a very strong complex is formed. Complementary oligo(dPu) does not form a complex under these conditions. No binding is observed for oligo(dPy) with non-homologous inserts as well as with vector plasmids without inserts. The complex is formed equally well with linear, nicked or superhelical DNA. The complex is not detected at pH greater than 6. Complex formation leads to very little, if any, unwinding of the duplex. The observed complex appears to be the Py.Pu.Py triplex consisting of TAT and CGC base-triads with protonated cytosines. Two-dimensional gel electrophoresis patterns show that the presence of homologous oligo(dPy) destabilizes the formation of the H form.

DNA↗
PubMed1987

The energetics of the B-Z transition in DNA.

The paper deals with the energetics of the transition to left-handed Z form in DNA with an arbitrary base sequence. There is a brief outline of the statistical-mechanical model of the B-Z transition allowing for three possible states of each base pair. The parameters of the model can be determined by comparing the theory with experimental data for the B-Z transition in inserts with given sequences in circular DNA. The model contains six energy parameters, most of which have been determined before. In order to find the remaining parameters of the model and test its adequacy, a number of oligonucleotide sequences were synthesized and inserted into the pUC 19 plasmid. Two-dimensional gel electrophoresis was used to determine the superhelical density at which the inserts adopt the Z form. A statistical-mechanical treatment of these data yielded a complete set of six energy parameters for the B-Z transition. The theoretical assumption that the free energy of Z-form pairs does not depend on the type of adjacent pairs proved to be in agreement with the experimental data.

Chemical Phenomena↗
PubMed1987

The ionic strength dependence of the cooperativity factor for DNA melting.

The melting temperature for the d(AT)24.d(AT)24 stretch, located inside the DNA helix and terminally, have been determined in a wide range of ionic strength values (0.01 - 1 M Na+). The cooperativity factor was calculated from the shifts in the melting temperature of the stretch due to its different boundary conditions. With the sodium concentration decreasing from 1 M to 0.01 M the cooperativity factor dropped by three orders of magnitude, its change being less marked at high than at low ionic strength.

DNA↗
PubMed1987

Structure of (dG)n.(dC)n under superhelical stress and acid pH.

We have recently shown that a (GA)n.(TC)n tract undergoes a sharp structural transition under superhelical stress (V.I. Lyamichev, S.M. Mirkin and M.D. Frank-Kamenetskii, J. Biomol. Struct. Dyn. 2,327 (1985]. Unlike the well studied transitions to the cruciform and to the Z form, this novel transition was strongly pH-dependent. We have found the (dG)n.(dC)n insert to undergo a pH-dependent structural transition similar to that of the (GA)n.(TC)n tract. These new data meet our earlier expectations and disagree with the data of D.E. Pulleyblank, D.B. Haniford and A.R. Morgan, Cell 42, 271 (1985). We conclude that a novel DNA structure (the H-form) is typical of homopurine-homopyrimidine mirror repeats (the H palindromes) under superhelical stress and/or acid pH. In the H-form the homopyrimidine strand forms a hairpin while half of the homopurine strand interacts with the hairpin forming a triplex, the other half of the homopurine strand being unstructured (V.I. Lyamichev, S.M. Mirkin and M.D. Frank-Kamenetskii, J. Biomol. Struct. Dyn. 2, 3, 667 (1986].

DNA↗
PubMed1987

[Transfer of "artificial transposons" constructed on the basis of insertion element IS1].

Terminal inverted repeats of the insertion element IS1 were synthesized chemically and plasmids containing these sequences flanking kanamycin-resistance gene in different combinations were constructed. Further incorporation of a whole-sized copy of the IS1 into such plasmids caused in some cases the autonomous transfer of Km-resistance from plasmid to bacteriophage lambda DNA. The transposition of the Km-resistance gene was only observed in those cases when the gene was enclosed between IS1 copy and one of the terminal repeats. The data obtained are discussed with regard to the evolution of bacterial transposons.

Bacteriophage lambda↗
PubMed1986

Structures of homopurine-homopyrimidine tract in superhelical DNA.

For homopurine-homopyrimidine tracts in superhelical DNA, we propose a structure involving Watson-Crick and Hoogsteen paired triple helixes, hairpin loops and unstructured domains. Topologically, the whole structure is equivalent to an open region. The proposed structure is consistent with available S1 cleavage, pH and alkylation data and energetics under superhelical stress; this new structure is a much more probable candidate than the one proposed by us recently (V.I. Lyamichev, S.M. Mirkin & M.D. Frank-Kamenetskii, J. Biomole. Str. Dyns 3, 327-338, 1985).

Base Sequence↗
PubMed1985

A pH-dependent structural transition in the homopurine-homopyrimidine tract in superhelical DNA.

We have inserted the 509-bp-long fragment of sea urchin P. miliaris histone gene spacer region into plasmid pUC19. The fragment contains the 60-bp-long homopurine-homopyrimidine tract that is known to be hypersensitive to the S1 endonuclease. Using two-dimensional gel electrophoresis we have observed a sharp structural transition in the insert with increasing DNA superhelicity. As in the cases of cruciform and Z form formation, the observed transition partly relaxes the superhelical stress. In contrast with the other two well documented transitions, the observed transition strongly depends on pH. At pH7 and above the transition occurs at negative superhelicities exceeding the physiological range (- sigma greater than 0.08). For pH6 the transition occurs at -sigma = 0.055, whereas for pH4.3 it takes place at -sigma = 0.001. A comprehensive analysis of the obtained data has made it possible to define the nature of the observed transition. We conclude that under superhelical stress or/and at low pH homopurinehomopyrimidine tracts adopt a novel spatial structure called the H form.

Animals↗
PubMed1985

[Development of bacteriophage Mu in E. coli gyrBts mutant strain].

The study deals with Mu growth in cells carrying a temperature-sensitive mutation in the gene of DNA gyrase B subunit. At a nonpermissive temperature the Mu growth is shown to be blocked in the host gyrB ts mutant both on infection and on prophage induction. Mu DNA does not get integrated in the host chromosome upon the infection of mutant cells, as demonstrated by DNA-DNA hybridization experiments. In the case of prophage induction in mutant cells, as opposed to the wild type cells early mRNA synthesis is practically fully inhibited while the total RNA synthesis is three times reduced after 20 min of induction. The transcription of phage DNA associated with the changed superhelicity of DNA in the cell.

Bacteriophage mu↗
PubMed1984

Native supercoiling of DNA: the effects of DNA gyrase and omega protein in E. coli.

This study deals with the effects of a temperature-sensitive (ts) mutation at the gene encoding the DNA gyrase B subunit (gyrBts) and a deletion of the top gene encoding the omega protein upon the superhelical density of the pAO3 plasmid in E. coli cells. The alteration of the DNA gyrase B subunit is shown to lead to a partial relaxation of DNA. On the other hand, the lack of omega protein due to the top gene deletion leads to an abnormally high degree of DNA supercoiling. In a double gyrBts delta top mutant the DNA supercoiling is greater than native at the permissive temperature, while under nonpermissive conditions a partial relaxation is observed. However, the pattern of DNA relaxation in the latter case is quite different from that in a single gyrBts mutant. The conclusion is that the native supercoiling of DNA in the cell is maintained through the counter-activities of DNA gyrase and the omega protein.

DNA Topoisomerases, Type I↗
PubMed1983

[Temperature-sensitive mutant Escherichia coli for the B-subunit of DNA-gyrase. The effect on replication and transcription].

A temperature sensitive mutant affecting the B-subunit of DNA gyrase was isolated. The mutation leads to the thermolability of DNA gyrase in vitro and to the plasmid DNA relaxation in vitro. The immediate stop of the increase in mutant culture titre has been observed under nonpermissive conditions. However, DNA synthesis does not cease, though its efficiency is reduced by a factor of three. The transcription rate is also reduced two - three times, but more quickly than the rate of replication. This would mean that the transcription apparatus is the first to react to the change of DNA supercoiling in the cell. This suggestion is supported by the facts that small amounts of rifampicin increase the viability of ts mutant cells under nonpermissive conditions and also by the previously obtained results concerning the change in sensitivity of RNA polymerase mutants to DNA gyrase inhibitors.

DNA Replication↗
PubMed1983

[Nature of the plasmids determining the unstable inheritance of transposon Tn9 in Escherichia coli K-12].

Unstable inheritance of transposon Tn9 in the Escherichia coli strain KS7201 had been connected with its integration into a certain bacterial chromosome site (attTn9A). However, the present work shows that the transposon is situated within an unstable plasmid in this strain. A possibility of such plasmid's formation, as a result of a deletion of a part of bacteriophage lambda DNA, is shown.

Bacteriophage lambda↗
PubMed1982

DNA replication and transcription in a temperature-sensitive mutant of E. coli with a defective DNA gyrase B subunit.

A temperature-sensitive mutant of E. coli with a defective DNA gyrase B subunit has been obtained. The mutation is expressed in the thermolability of DNA gyrase in vitro and in DNA relaxation in vivo. DNA replication in the mutant does not stop under non-permissive conditions; its rate gradually falls by a factor of 2 to 3. The transcription rate also drops by a factor of 2 to 3, but before replication. Small concentrations of rifampicin, an inhibitor of bacterial RNA polymerase, make for a partial survival of the mutant cells under non-permissive conditions. The results suggest the conclusion that DNA supercoiling is mainly required to ensure the optimum transcription level in the cell.

Cell Division↗
PubMed1982

Changed properties of the A subunit in DNA gyrase with a B subunit mutation.

To investigate the interaction of subunits A and B of DNA gyrase during DNA supercoiling, a Cour mutant of Escherichia coli was obtained and the effect of nalidixic acid on the supercoiling of DNA by wild-type and mutant enzymes was assayed. The enzyme of the Cour strain proved to be more sensitive to nalidixic acid than the wild-type DNA gyrase. Hence the mutation affecting the B subunit can also change the properties of the A subunit, which fact suggests that the two subunits of DNA gyrase are in contact during DNA supercoiling.

DNA Topoisomerases, Type II↗
PubMed1979

DNA supercoiling and transcription in Escherichia coli: influence of RNA polymerase mutations.

Coumermycin A1, a specific inhibitor of DNA gyrase, differentially changes the spectrum of proteins synthesized in wild type E. coli cells but has no effect on the protein spectrum in mutant cells with coumermycin-resistant DNA gyrase. The rpoB265 mutation affecting RNA polymerase decreases the coumermycin A1-sensitivity of bacteria while the rpoC3 mutation increases it. The interaction of wild type and mutant RpoB265 RNA polymerases with ColEl plasmid DNA in vitro is differently affected by DNA supercoiling. No such differences are observed in the case of RpoC3 RNA polymerase. The results suggest that template supercoiling may have a substantial effect on transcription in vivo, an effect which, in some cases, depends on the properties of RNA polymerase.

Aminocoumarins↗
PubMedDate not supplied

DNA H form requires a homopurine-homopyrimidine mirror repeat.

Regular homopurine-homopyrimidine tracts, (dG-dA)n(dT-dC)n and (dG)n(dC)n, undergo a superhelix-induced, strongly pH-dependent, structural transition into a novel DNA conformation, the H form. We have suggested that the H form can arise in any homopurine-homopyrimidine mirror repeat (H palindrome). We have now tested this prediction using a tailored series of plasmids carrying the inserts AAGGGAGAAXGGGGTATAGGGGYAAGAGGGAA, where X and Y may be either A or G, and subject them to two-dimensional gel electrophoresis. In support of our hypothesis, the inserts exhibited facile transitions into the H form for X = Y = G, or X = Y = A, whereas the transition was much more difficult or impossible for the two non-palindromes (X = A, Y = G or X = G, Y = A). We present evidence that the H form is the structural basis for S1-nuclease hypersensitivity.

Animals↗
PubMedDate not supplied

[Expansion of trinucleotide repeats].

This review describes a novel type of genome instability, expansion of trinucleotide repeats. Originally discovered in 1991 upon cloning the gene responsible for the fragile X syndrome, it appeared to be a general phenomenon responsible for a growing number of human neurological disorders. Besides apparent medical importance, the discovery of trinucleotide repeat expansion unraveled a fundamental problem of human genetics: a non-Mendelian type of inheritance called anticipation. Understanding the mechanisms of repeat expansion and the molecular pathways leading from these expansions to human diseases became a formidable task for modern biology and one of its spectacular achievements. Here we discuss the major breakthroughs in this field made during the last decade with an emphasis on molecular models of repeat expansion.

Animals↗