Identification of a cis-acting element involved in the regulation of BACE1 mRNA alternative splicing.
beta-site APP cleaving enzyme 1 (BACE1) is the transmembrane aspartyl protease that catalyzes the first cleavage step during proteolysis of the beta-amyloid precursor protein, a process involved in the pathogenesis of Alzheimer disease. BACE1 pre-mRNA undergoes complex alternative splicing, and cis-acting elements important for its regulation have not been identified. We constructed and compared several BACE1 minigenes and found that BACE1 sequence from exon 3 through exon 5 was required for minigenes to undergo correct splicing. Minigene splicing was validated by showing specific splicing inhibition upon splice site mutation. Furthermore, we showed that mutation of the minigene at a predicted exonic splicing enhancer in exon 4 of BACE1 increased exon 4 skipping. Therefore, we have for the first time found evidence of a regulatory site involved in BACE1 alternative splicing, and these data indicate that minor sequence changes can dramatically alter BACE1 alternative splicing.
Mowrer
Karen R
KR
Center for Neurologic Diseases, Brigham and Women's Hospital and Harvard Medical School, Boston, Massachusetts, USA.
Wolfe
Michael S
MS
D017398:Alternative Splicing / Q000235:genetics*; D053829:Amyloid Precursor Protein Secretases / Q000096:biosynthesis / Q000235:genetics*; D016282:Aspartic Acid Endopeptidases / Q000096:biosynthesis / Q000235:genetics*; D002460:Cell Line; D005091:Exons / Q000235:genetics; D006801:Humans; D016296:Mutagenesis / Q000187:drug effects; D009154:Mutation / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D011487:Protein Conformation; D012333:RNA, Messenger / Q000096:biosynthesis* / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D012333:RNA, Messenger; D053829:Amyloid Precursor Protein Secretases; D016282:Aspartic Acid Endopeptidases; C509280:BACE1 protein, human