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Nucleic Acids Research, 1989, Vol. 17, No. 10 3795-3810
© 1989

CHEMISTRY

Direct evidence for the presence of left-handed conformation in a supramolecular assembly of polynucleotides

T.J. Thomas and Thresia Thomas1

1Environmental and Community Medicine, UMDNJ-Robert Wood Johnson Medical School New Brunswick, NJ 08903, USA Departments of Medicine UMDNJ-Robert Wood Johnson Medical School New Brunswick, NJ 08903, USA

Received April 10, 1989. Revised April 20, 1989. Accepted April 20, 1989.

Hexammine cobalt(III) chloride (Co(NH3)63+) provokes a B-DNA -> Z-DNA -> Â{Psi}-DNA conformational transition in poly(dG-dC).poly(dG-dC) and poly(dG-m5dC).poly(dG-m5dC). The circular dichroism spectrum of S'-DNA is characterized by a manyfold increase of positive ellipticity in the range of 300-225 nm and the complete absence of a negative peak. In order to ascertain the helical handedness of >P-DNA, we used a recently developed enzyme immunoassay technique. This method consisted of treating the polynucleotides with Co(NH3)63+ to convert them to the Z- or y-DNA forms and immobilizing these conformations on a microtiter plate. The plates were subsequently treated with a monoclonal anti-Z-DNA antibody Z22, alkaline phosphatase conjugated, affinity purified immunoglobulins, and the phosphatase substrate. The enzyme-substrate reaction was monitored by reading the absorbance at 405 nm with a microplate autoreader. The monoclonal anti-Z-DNA antibody had no reactivity to the B-DNA form, but bound strongly to both the Z- and {Psi}-DNA forms, showing that Co(NH3)63+-induced ÂF-DNA form of the polynucleotides exists in the left-handed Z-DNA conformation.


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