The B form to Z form transition of poly(dG-m5dC) is sensitive to neutral solutes through an osmotic stress.

Several neutral solutes, ranging in size from methanol to a tetrasaccharide, stachyose, are shown to stabilize the left-handed Z form of the methylated polynucleotide poly(dG-m5dC). The action of these solutes is consistent with an osmotic stress, that is, with their effect on water chemical potentials coupled to a difference in the number of associated water molecules between the B and Z conformations. The apparent difference in hydration between the two forms is, however, dependent on the particular solute used to probe the reaction. The effect of solutes is not consistent either with a direct binding of solute or with an indirect effect on electrostatics or ion binding through changes in the solution dielectric constant. The interplay of NaCl and neutral solute in modulating the B-Z transition suggests that salt also could be stabilizing the Z form through an osmotic stress.

Hydrophobic moieties in cations, anions, and alcohols promote the B-to-Z transition in poly[d(G-C)] and poly[d(G-m5C)].

Previous results from our laboratory showed that an increase in the nonpolar alkyl chain length in tetraalkylammonium cations is accompanied by greater efficiency in driving the B-to-Z transition. Analogous effects are observed when the hydrocarbon portion of carboxylate anions and alcohols is increased in size. The more hydrophobic species have a greater ordering effect on the aqueous solvent and promote formation of the less hydrated Z-DNA conformer.

The B-DNA to Z-DNA transition in alkali and tetraalkylammonium salts correlated with cation effects on solvent structure.

The B-to-Z transition in DNA with alternating purine and pyrimidine sequences is driven by high concentrations of monovalent cations. In addition to screening of phosphate repulsion, an important factor may be the influence of ions on DNA hydration. The relative efficiencies of tetraalkylammonium and alkali metal salts in promoting the formation of Z-DNA, monitored by CD, in poly[d(G-C)] and poly[d(G-m5C)] was correlated with the degree to which the cations tie up water molecules in ordered arrangements and therefore decrease the availability of the solvent to hydrate DNA.

Evolutionary sequence divergence within repeated DNA families of higher plant genomes. II. Analysis of thermal denaturation.

An assay based on derivative analysis of thermal denaturation (melting) behavior of reassociated DNA was developed in an attempt to characterize the sequence relationships in repeated DNA families according to the homogeneous or heterogeneous models of Bendich and Anderson (1977). The validity of the technique was confirmed by the use of deaminated Escherichia coli DNA models for repetitive families. The melting data for DNA reassociated at two different temperatures provided strong evidence that Pisum sativum repeated families are mostly heterogeneous, while homogeneous families predominate in Vigna radiata. These findings, together with other differences between the two genomes, suggest that the rate of sequence amplification has been higher in the evolutionary history of Pisum DNA. A general trend seems to exist for high amplification rates in large, highly repetitive plant genomes such as Pisum and lower rates in smaller plant genomes such as Vigna, as well as in the generally smaller, less repetitive genomes of most animal species.

Evolutionary sequence divergence within repeated DNA families of higher plant genomes. I. Analysis of reassociation kinetics.

The higher proportion of repeated DNA sequences in the garden pea (Pisum sativum) than in the mung bean (Vigna radiata), as well as other differences between these legume genomes, are consistent with a higher rate of sequence amplification in the former. This hypothesis leads to a prediction that repeated sequence families in Pisum are mostly heterogeneous, as defined by Bendich and Anderson (1977), while Vigna families are homogeneous. An assay developed by these authors to distinguish between the two types of families, by comparison of reassociation rates at different temperatures, was utilized. The results for Vigna defied the predictions of the assay for either homogeneous or heterogeneous model. Evaluation of the kinetic data in light of the great diversity of repeated family copy numbers in both genomes enabled an interpretation of the results as consistent with heterogeneous families in Pisum and homogeneous families in Vigna. These tentative conclusions were supported by the results of a thermal denaturation (melting) assay described in the accompanying paper.