Base extrusion is a major structural feature at the junction between B- and Z-DNA (the B–Z junction) where a base pair is broken, and the two bases are extruded from the double helix. Despite the demonstration of base extrusion at the B–Z junction, it is not clear whether a similar base extrusion occurs at other types of junctions involving the left-handed Z conformation. Here, we investigate structural changes of bases at three Z-form junctions: DNA B–Z and Z–Z and RNA A–Z junctions. By monitoring fluorescently labeled duplex nucleic acids using 2-aminopurines at various positions relative to the junction point, we show that base extrusion occurs not only at the DNA B–Z junction, but also at the RNA A–Z and DNA Z–Z junctions. Our data suggest that base extrusion is a general feature of Z-form nucleic-acid junctions.Korean Science and Engineering Foundation (grant NRL-2006-02287)Korea (South). Ministry of Science and Technology (21C Frontier Functional Proteomics Program FPR08B2-270)Korean Science and Engineering Foundation (Ubiquitome Research Program M105 33010001-05N3301-00100)Korea Research Foundation (MOEHRD, Basic Research Promotion Fund; KRF-2005-070-C00078