Expression and regulation of a low-density lipoprotein receptor exon 12 splice variant.
As low-density lipoprotein receptor (LDLR) contributes to cholesterol and amyloid beta homeostasis, insights into LDLR regulation may facilitate our understanding of cardiovascular disease and Alzheimer's disease. Previously, we identified LDLR isoforms that lacked exon 12 or exons 11-12 and that are predicted to encode soluble, dominant negative, LDLR. Moreover, these isoforms were associated with rs688, an exon 12 polymorphism that was associated with LDL-cholesterol and Alzheimer's disease risk. In this study, we present evidence that although the truncated LDLR isoforms are translated in vitro, they represent < 0.1% of CSF proteins. As these LDLR isoforms likely represent a loss of mRNA-encoding functional LDLR, we then focused upon identifying intron-exon boundary and exonic splicing enhancer elements critical to splicing. Exon 12 inclusion is enhanced by altering the 5' splice site in intron 12 towards a consensus splice donor sequence, consistent with its being a weak 5' splice site. Additionally, of the nine evolutionarily conserved putative splicing enhancer regions within exon 12, two regions that flank rs688 were critical to exon 12 inclusion. Overall, these results suggest that LDLR splice variants represent a loss of mRNA encoding functional LDLR and provide insights into the regulatory elements critical for LDLR exon 12 splicing.
Ling
I-Fang
IF
Department of Physiology, Sanders-Brown Center on Aging, University of Kentucky, Lexington, Kentucky 40536-0230, USA.
Gopalraj
Rangaraj K
RK
Simpson
James F
JF
Estus
Steven
S
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D017124:Conserved Sequence; D005091:Exons / Q000235:genetics; D006801:Humans; D007438:Introns / Q000235:genetics; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009690:Nucleic Acid Conformation; D010957:Plasmids / Q000235:genetics; D011110:Polymorphism, Genetic / Q000235:genetics; D020033:Protein Isoforms / Q000235:genetics*; D012313:RNA / Q000737:chemistry / Q000235:genetics; D011973:Receptors, LDL / Q000096:biosynthesis / Q000235:genetics* / Q000502:physiology*; D050436:Regulatory Elements, Transcriptional / Q000235:genetics / Q000502:physiology; D013045:Species Specificity
D020033:Protein Isoforms; D011973:Receptors, LDL; D012313:RNA
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