In this report we investigate the expression and relationship of transgelin (Tg), a transformation and shape-change sensitive actin gelling protein found in fibroblasts and smooth muscle [Shapland et al., 1988: J. Cell. Biol. 107:153-161; Shapland et al., 1993: J. Cell. Biol. 121:1065-1073], to SM22alpha, a smooth muscle protein of unknown function [Lees-Millar et al., 1987: J. Biol. Chem. 262:2988-2993; Solway et al., 1995: J. Biol. Chem. 270:13460-13469]. To clarify their relationship we have cloned and sequenced the cDNA encoding Tg from cultures of rat embryo fibroblasts. The sequences of fibroblast Tg and the smooth muscle isoform SM22 are identical [Prinjha et al., 1994: Cell Motil. Cytoskeleton 28:243-255; Shanahan et al., 1993: Circ. Res. 73:193-204; Solway et al., 1995]. These data, coupled with our immunoblot analysis and previous observations [Shapland et al., 1988; Shapland et al., 1993], demonstrate that Tg expression is not restricted to smooth muscle since this protein is also present in normal mesenchymal cells. However, we also show that Tg, although present in secondary cultures of mouse and rat embryo fibroblasts, is absent in many apparently normal fibroblast cell lines. Tg down-regulation may therefore be an early and sensitive marker for the onset of transformation. A functional role for Tg is unlikely to directly involve Ca2+ since it neither contains a functional EF hand nor binds 45Ca2+.