Galectin-3: differential accumulation of distinct mRNAs in serum-stimulated mouse 3T3 fibroblasts

The murine Galectin-3 gene spans approximately 12 kb of DNA and contains six exons, with the translation initiation codon located in exon II. On the basis of restriction mapping and sequence analysis of the DNA upstream of exon II, primer extension assays, rapid amplification of cDNA ends, and ribonuclease protection assays were designed and carried out to determine the initiation site of transcription and the sequence of exon I. The results revealed at least two transcription initiation sites (alpha and delta), each of which appears to be specifically associated with the use of alternative donor splice sites, resulting in distinct mRNA species. Type I message initiates at transcription start site delta, uses splice donor site No. 2, retaining a 27 bp sequence, whereas type II message initiates at transcription start site alpha, uses splice donor site No. 1, resulting in the loss of the 27 bp sequence. Primer extension assays carried out with mRNA isolated from 3T3 fibroblasts at various times after serum stimulation indicate that while the type II message varies in level only a little over the first 20 h, there is dramatic accumulation of the type I message, which peaks at 16 h post mitogen addition.