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Nucleic Acids Research, 1989, Vol. 17, No. 20 8257-8271
© 1989

MOLECULAR BIOLOGY

Magnesium-dependent supercoiling-induced transition in (dG)n·(dC)n stretches and formation of a new G-structure by (dG)n strand

I.G. Panyutin, O.I. Kovalsky and E.I. Budowsky

N.D.Zelinsky Institute of Organic Chemistry, USSR Academy of Sciences 47 Leninsky pr., Moscow 117913, USSR

Received August 1, 1989. Revised September 25, 1989. Accepted September 25, 1989.

Plasmids containing (dG)27· (dC)27 inserts (pPG27), (dG)37· (dC)37 inserts (pPG37), and (dG)24C(dG)21 (dC)24 G(dC)21 inserts (pPG460) were oonstruoted for the study of structural Transitions within (dG)n ·(dC)n stretohes. Two-dimensional gel eleotrophoresis has shown that a Mg2+-dependent supercoiling-induced structural transition takes place at pH 8 in plasmid pPG46C. The transition occurs at –0=0.06 and involves a supercoiling release corresponding to 5 superhelical turns.

After denaturation of the restriction fragments containing (dG)n ·(dC)n inserts, the strands do not renature completely and the (dG) -containing strand migrates in PAGE much faster than the (dC)n -containing one. Chemical modification experiments with the (dG)n-strand have revealed the periodic nature of the protection of guanines against dimethyl sulfate methylation. The (dG) strand in the presence of Mg2+ forms complexes with the complementary (dC)n strand, which differ from the native duplex in mobility. We believe these effects to be due to the formation of an intrastrand structure within the (dG)n strand stabilized by G·G interactions, (we called it G-structure), which in the presence of Mg2+ forms an interstrand complex. with the (dC)n strand.


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