Linear dichroism studies of binding site structures in solution: Complexes between DNA and basic arylmethane dyes

The interaction between B-form DNA and twelve cationic triaryl-methane dyes was studied with respect lo optical properties and stabilities, using linear dichroism (LD) and aqueous two-phase partition techniques. Monovalent dyes derived from crystal violet as a rule form a single strong complex (K₁ ca 10⁵ M^?1; site density per nucleotide base n₁ ca 0.1 at 0.1M ionic strength) in which the plane of the dye is at an angle of less than 50° to the local DNA helix axis. The complex with fuchsin is weaker (10⁴M^?1) but can be explained by a similar orientation. For some of the dyes (those with pseudo-C_2v symmetry) XXXre angular orientations of two molecule-fixed axes can be obtained. For the divalent methyl green a second complex appears to be formed at low ionic strength. Methyl green (and to some extent 2-thiophene green and malachite green) show exciton splitting in the LD spectrum and circular dichroism assignable to exciton coupling between transition dipoles roughly parallel to the helical strands, indicating a dye-dye interaction. Tne optical data, supported by fitting experiments with space-filling models, suggests a general structure for the binding site. The dye is not intercalated but is bound to exposed hydrophobic regions in the major groove. The ligand is in part (the charged amino groups) in contact with the phosphoribose chain but its main surface lies against the hydrophobic base-pair stack. For a diphenylmethane dye, Michler's hydrol blue, a perpendicular orientation was observed, possibly due to intercaiation.     

A DESEEDED AVENA TEST METHOD FOR SMALL AMOUNTS OF AUXIN AND AUXIN PRECURSORS

The main results presented in this article may be summarized as follows: 1. A test method with deseeded Avena seedlings for small concentrations of auxin and precursors of auxin has been described. 2. This method makes possible quantitative determinations of about ten times as low concentrations of hormone as can be obtained with the standard method, (a) Through an increase in the time of the test, so that nearly all the hormone applied can be utilized. (b) Through an increase in sensitivity of deseeded plants to unilaterally applied small concentrations of hormone. 3. The effect of deseeding in relation to curvature growth is primarily the prevention of auxin regeneration through the removal of the material for auxin synthesis, and in addition the prevention of physiological aging. 4. The mechanism of auxin synthesis in the tip of the coleoptile and the mechanism of auxin regeneration in the new physiological tip have been shown to be identical. 5. The application of the deseeded method is illustrated by determinations of auxin in primary leaves and coleoptile sections of Avena seedlings. 6. The deseeded method has been used as a test method for precursors of auxin obtainable from the coleoptile and from other sources. The method further makes possible a distinction between auxins and these substances which may become activated by the plant. 7. Evidence for the existence of a precursor of auxin in the plant is given (a) indirectly by determinations of the decrease in auxin synthesis in deseeded plants. (b) Directly by its isolation from the plant. 8. Precursors of hetero-auxin are demonstrated; their chemical nature and activation are briefly considered.     

Cytokinin antagonists: Synthesis and physiological effects of 7-Substituted 3-Methylpyrazolo[4,3-d]Pyrimidines

A series of sixteen pyrazolo[4,3-d]pyrimidine derivatives has been synthesized and tested for cytokinin antagonist activity in the tobacco bioassay