The human vinculin gene contains 22 exons ranging in size from 71 base pairs (bp) to 303 bp (average 155 bp) with the exception of exon 22 which contains 144 bp of coding sequence and 1848 bp of 3'-untranslated sequence including two polyadenylation signals. There is a limited correlation between exon boundaries and functional domains within the vinculin molecule. The talin-binding domain in vinculin spans residues 1-258, and the first 6 exons encode residues 1-261. Similarly, the predicted boundaries of the central repeat domain (residues 259-589) are close to the boundaries of exons 7 and 12. Analysis of vinculin mRNAs in human uterus showed that alternative splicing of the gene is limited to exon 19, which encodes the 68 amino acids included in the muscle-specific isoform called metavinculin. We have determined 1.1 kilobases of sequence 5' of the transcription start site. The vinculin promoter lacks a TATA box, but does contain six Sp1 sites, and a CArG box at position -262 which forms the core of the serum response element found in immediate-early response genes. Expression of a vinculin promoter/CAT construct is serum-inducible in NIH3T3 cells demonstrating that the promoter does contain a functional serum response element.