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 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+).

[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.

[Effect of superhelical DNA on the transcription of cloned genes of the T4 phage].

The effect of DNA superhelicity on the transcription of T4 DNA fragments containing early genes uvs W, Y and late genes 25-29 was studied. RNA polymerase transcribes both early and late phage genes within the supercoiled recombinant plasmid. Late genes relative transcription increases essentially when T4-modified RNA polymerase is used.

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.

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.

A structural transition in d(AT)n.d(AT)n inserts within superhelical DNA.

We have constructed plasmids carrying d(AT)n.d(AT)n inserts of different lengths. Two-dimensional gel electrophoresis patterns show that an increase in the negative superhelicity of these DNAs brings about a structural transition within the inserts, resulting in a reduction of the superhelical stress.

[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.

The absence of cruciform structures from pAO3 plasmid DNA in vivo.

We extracted pAO3 plasmid DNA from E. coli cells, having "frozen" the transition between cruciform and double-helical conformations in DNA. The characteristic feature of the DNA isolation procedure is that all steps were carried out at temperature between 0 and 4 C and no phenol deproteinization was used, since it has been discovered that phenol destabilizes cruciform structures in pAO3 DNA.

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.

[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.

[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.

[Interactions of subunits of DNA gyrase].

Interaction of DNA gyrase A- and B-subunits during the process of DNA supercoiling was studied. For this purpose a E. coli Cour-1 mutant resistant to coumermycin and containing a mutation in the B-subunit of DNA gyrase was isolated and the influence of the DNA gyrase A-subunit specific inhibitor-nalidixic acid-on DNA supercoiling by wild-type and mutant enzymes was investigated.

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.

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.

Empirical assessment of competency to consent to psychiatric hospitalization.

The authors used an interview questionnaire to assess competency to consent to voluntary admission in a group of 50 newly admitted psychiatric patients. Using a number of alternative definitions of competency with this sample of patients tested shortly after admission, they found that a majority of patients appeared to have severe impairment of competency. These results suggest limitations in the applicability of the legal model of informed consent to the psychiatric setting.

[Influence of DNA superhelicity on the major genetic processes in prokaryotes].

Some properties of superhelical DNA molecules and enzymes (topoisomerases) changing DNA superhelical density are discussed. Data on the effect of DNA supercoiling on replication, transcription, and recombination, and the participation of topoisomerases in these processes are presented.

[Effect of DNA supercoiling on transcription performed by normal and mutant Escherichia coli RNA-polymerases].

A specific DNA-gyrase, inhibitor, coumermycin A1, causes differential changes in the spectrum of proteins synthesized in E. coli wild types cells, while protein spectrum in the mutant cells with coumermycin-resistant DNA-gyrase is left unaffected. The mutation of RNA-polymerase RpoB265 lowers the sensitivity of bacteria to coumermycin A1, whereas the RpoC3 enhances it.

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.