Analysis of the human integrin alpha11 gene (ITGA11) and its promoter

Integrin alpha11beta1 is a collagen receptor which is expressed in a subset of mesenchymally-derived tissues during embryogenesis. Based on available human chromosome 15-derived sequences and genomic PCR, the complete exon structure of ITGA11, including the proximal promoter, was assembled into 30 exons. The inserted region (encoding amino acids 804-826) distinguishing alpha11 from other integrin alpha chains, was placed in the very beginning of exon 20. PCR data failed to show alternative splicing of RNA transcribed from this region. Using the oligo-capping technique a major transcription start site was mapped 30 nucleotides upstream of the translation start and identified as an abbreviated initiator sequence. Promoter sequence analysis in silico suggested the presence of multiple binding sites for transcription factors in the region upstream of the transcription start. 3 kb of the 5' flanking sequence was isolated and used to generate luciferase promoter constructs. In the fibrosarcoma cell line HT1080 a core promoter [nt (-)127-(+)25], a potential silencer region [nt (-)400-(-)127] and a potential enhancer region [nt (-)1519-(-)400], were identified as being important for alpha11 transcription in mesenchymal cells. Furthermore, studies of the promoter region will provide valuable information regarding the molecular mechanisms underlying the cell- and tissue- specific expression pattern of ITGA11.