Phylogenetic relationships among plants, animals, and fungi were examined by using sequences from 25 proteins. Four insertions/deletions were found that are shared by two of the three taxonomic groups in question, and all four are uniquely shared by animals and fungi relative to plants, protists, and bacteria. These include a 12-amino acid insertion in translation elongation factor 1 alpha and three small gaps in enolase. Maximum-parsimony trees were constructed from published data for four of the most broadly sequenced of the 25 proteins, actin, alpha-tubulin, beta-tubulin, and elongation factor 1 alpha, with the latter supplemented by three new outgroup sequences. All four proteins place animals and fungi together as a monophyletic group to the exclusion of plants and a broad diversity of protists. In all cases, bootstrap analyses show no support for either an animal-plant or fungal-plant clade. This congruence among multiple lines of evidence strongly suggests, in contrast to traditional and current classification, that animals and fungi are sister groups while plants constitute an independent evolutionary lineage.