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J. Biochem, 1996, Vol. 119, No. 4 805-810
© 1996 Japanese Biochemical Society

research-article

Formation of a Triple-Stranded DNA between d(GGA: TCC) Repeats and d(GGA) Repeat Oligonucleotides1

Yukio Mishima2, Takeshi Suda and Ryo Kominami

Department of Biochemistry, Niigata University School of Medicine 1-757 Asahimachi-dori, Niigata 951

2To whom correspondence should be addressed. Fax: + 81-25-223-0237

Incubation of the 162-bp duplex containing d(GGA:TCC)11 repeats with d(GGA),, and d(GGT)11 oligonucleotides, but not with d(TCC)11, gave a mobility-shifted band in poly-acrylamide gel electrophoresis, suggestive of the triplex formation. The two conformers, however, showed difference in binding affinity and structure. The complex formed with d(GGT)11 exhibited patterns protected from DNase I digestion and from modifications with osmium tetroxide and dimethyl sulfate. The patterns were conformed to a proposed triplex model. In contrast, the complex with d(GGA)11 was sensitive to DNase I and to osmium tetroxide, which is reactive to pyrimidine bases of single-stranded DNA. These results suggest that the d(GGA:TCC)11 and d(GGA)11 triple-stranded complex forms a D-loop-like structure. By considering the ability of d(GGA)11 oligonucleotides to form a parallel homoduplex, the structure may be one in which the GGA-strand of the duplex pairs with d(GGA)11 in parallel, with the TCC-strand being looped out.

1 This work was supported in part by Grants-in-Aid for Scientific Research from the Ministry of Education, Science and Culture of Japan.


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