Schizophyllan is an extracellular polysaccharide consisting of a beta-1,3-D-glucan main chain and exists as a triple helix in water and as a single chain in dimethyl sulfoxide (DMSO). When the single chain of schizophyllan (s-SPG) was mixed with poly(C), poly(A), poly(dA), or poly(dT), they form a macromolecular complex. On the other hand, poly(G), poly(U), poly(I), poly(dG), and poly(dC) do not. This nucleotide specificity evidences that the hydrogen bonds are essential to form the complex, because the former nucleotides have an unoccupied hydrogen-bonding site and the latter ones use the hydrogen-bonding sites in the intramolecular aggregation (i.e., such as the G quartet for poly(G) and poly(dG) and the U hairpin for poly(U)). The hypochromic effect and the increment in the circular dichroism (CD) intensity are observed in accordance with the complex formation. These facts indicate that the base stacking is enhanced in the complex. The solvent-composition (DMSO/water) dependence demonstrates that the hydrophobic interaction is important to form the complex as well as the hydrogen-bonding interaction. With increasing temperature the complex dissociates cooperatively and the melting curve enables the thermodynamic parameters to be evaluated (delta H = -60 to 70 kcal mol-1 and delta S = -150 to 200 cal mol-1 K-1). These values are comparable with those for double helix DNA. Namely, the complex can be characterized by enhancement of the base stacking, cooperative dissociation, the similar thermodynamic parameters to DNA, and combination of the hydrogen-bonding and hydrophobic interactions to form the higher-order structure. These facts surprisingly coincide with characters of the double helix of DNA. In other words, the s-SPG molecule behaves as if it were a complementary polynucleotide chain for the corresponding polynucleotide. Furthermore, stoichiometric study suggested that the complex structure is a triple helix consisting of two s-SPG and one poly(C) or poly(A) chains.