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Nucleic acid junctions are stable analogs of branched DNA structures which occur transiently in living systems. We show here that junctions which contain three double helical arms can be enzymatically oligomerized, using conventional sticky-ended ligation procedures, to create larger complexes. The products consist of a series of linked junctions separated by 20 base pairs. Junction dimers are formed that have free termini only, whereas trimers and larger species are found to be both unclosed and cyclized. The formation of a series of macrocyclic products which, surprisingly, begins with trimers and tetramers indicates that this junction is flexible about a bending axis, and perhaps twist-wise as well. We have obtained the same results from three different 3-arm junctions, two in which the junction is flanked by a 3 Watson-Crick base pairs, and one in which a G-G base pair flanks the junction.

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An immobile nucleic acid junction composed of four dodecanucleotides has been designed according to principles of minimum symmetry aided by equilibrium calculations, and has been synthesized by automated phosphotriester techniques. We can demonstrate its tetrameric character and its 1:1:1:1 stoichiometry by gel electrophoresis. Thermal denaturation monitored by ultraviolet hyperchromism indicates that the complex is stable relative to its component arms. High resolution NMR spectroscopy suggests that this junction exists in more than one conformer at room temperature. The data from this junction are compared with the data from a similar junction composed of four hexadecanucleotides.

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