High-mobility group I (HMGI) proteins are architectural transcription factors expressed predominantly during embryonic development. Their genetic loci are the most frequent targets of chromosomal rearrangements in uterine leiomyomas and other benign tumors. It was therefore suggested that both HMGI genes are involved in the neoplastic transformation of benign tumors. By Western analysis we found that 16 of 33 uterine leiomyomas expressed high levels of HMGI-C or HMGI(Y) proteins, whereas they were not detected in the corresponding myometrium. Immunohistochemistry demonstrated that the expression of HMGI-C is restricted to leiomyoma smooth muscle cells but is not expressed in vascular smooth muscle cells or the connective tissue of the tumor. Northern blotting confirmed the protein expression data for HMGI-C, whereas HMGI(Y) mRNA and protein levels did not correlate, suggesting that posttranscriptional mechanisms are involved in the regulation of HMGI(Y) expression. Three of the uterine leiomyomas analyzed expressed HMGI-C gene products with altered molecular weight. Two of them were proved to consist of the entire DNA-binding domain but lacked sequences of the C-terminal acidic tail. Conversely, other tumors expressed HMGI-C or HMGI(Y) genes that were not affected by mutations of the coding region. Thus we identified uterine leiomyomas that expressed mutated HMGI-C, whereas other uterine leiomyomas expressed wild-type HMGI-C or HMGI(Y). On the basis of our data we assume that the enhanced expression of functionally active HMGI proteins, whether they are wild-type or not, is important for the pathogenesis of uterine leiomyomas.