NDLI: Formation of DNA triplexes accounts for arrests of DNA synthesis at d(TC)n and d(GA)n tracts.

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Formation of DNA triplexes accounts for arrests of DNA synthesis at d(TC)n and d(GA)n tracts.

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Formation of DNA triplexes accounts for arrests of DNA synthesis at d(TC)n and d(GA)n tracts.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Baran, N. Lapidot, A. Manor, H.
Description	Author Affiliation: Baran N ( Department of Biology, Technion-Israel Institute of Technology, Haifa, Israel.);
Abstract	To study the mechanism of arrest of DNA synthesis at d(TC)n and d(GA)n sequences, single-stranded DNA molecules including d(TC)27 or d(TC)31 tracts or a d(GA)27 tract were used as templates for in vitro assays of complementary DNA synthesis performed by extension of a primer with the Klenow polymerase or the Taq polymerase (Thermus aquaticus DNA polymerase). Electrophoresis of the products revealed that arrests occurred around the middle of these tracts. The arrests in the d(TC)n sequences were eliminated when dATP or dGTP was replaced with the analogue 7-deaza dATP or 7-deaza dGTP, respectively, or when the templates were preincubated with the Escherichia coli single-strand binding protein (SSB). Preincubation of the template including a d(GA)27 tract with SSB has also eliminated the arrests at this sequence. Furthermore, arrests did not occur at d[G(7-deaza A)]27 or d[(7-deaza G)A]27 tracts when molecules including such tracts were used as templates. These results are compatible with the notion that the arrests were caused by formation of d(TC)i.d(GA)i.d(TC)i and d(GA)i.d(GA)i.d(TC)i triplexes, in which the bases in the uncopied portions of the d(TC)n tracts, or of the d(GA)27 tract, and the purine bases in the newly synthesized d(TC)i.d(GA)i duplexes were bound by hydrogen bonds. In the assays performed with the Taq polymerase, the pH dependence (in the range of 6.0-9.0) and the temperature dependence of the arrests were determined. As the pH was lowered, the arrests in the d(TC)27 tract were enhanced, in line with the expected properties of d(TC)i.d(GA)i.d(TC)i triplexes. The arrests in the d(GA)27 tract were enhanced by an increase in the pH. At pH 7.2 the arrests in the d(GA)27 tract persisted up to 80 degrees C, whereas the arrests in the d(TC)27 tract were eliminated at 50 degrees C; these results presumably reflect the relative stabilities of the two triplexes mentioned above at this physiological pH value and could be biologically significant.
ISSN	00278424
e-ISSN	10916490
Journal	Proceedings of the National Academy of Sciences of the United States of America
Issue Number	2
Volume Number	88
Language	English
Publisher	National Academy of Sciences
Publisher Date	1991-02-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	DNA Replication DNA, Single-Stranded Biosynthesis DNA-Directed DNA Polymerase Metabolism Polydeoxyribonucleotides Adenine Analogs & Derivatives DNA-Binding Proteins Escherichia Coli Guanine Hydrogen Bonding Kinetics Models, Structural Molecular Sequence Data Nucleic Acid Conformation Polymerase Chain Reaction Templates, Genetic Thermodynamics Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Multidisciplinary
Content Type	Text
Resource Type	Article
Subject	Multidisciplinary

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