Identification of a CpG island in the human LRP-2 gene and analysis of its methylation status in parathyroid adenomas.
Methylation of CpG residues in mammalian genomes is a mechanism of vital importance for many cellular functions, which all relate to gene expression. In this study we describe the identification of a CpG island in the 5'-region of the gene encoding human megalin/LRP-2, a receptor capable of binding multiple ligands, which is involved in the regulation of calcium metabolism. Southern blot analysis and genomic bisulfite sequencing revealed that the CpG island is methylated in a non-expressing cell line, largely unmethylated in an expressing cell line and unmethylated in human parathyroid tissue. In addition, we show that artificial methylation of LRP-2 promoter reporter plasmids leads to strong transcriptional repression, in vitro as well as in transfected cells. No evidence for aberrant LRP-2 gene methylation in parathyroid adenomas, in which the LRP-2 protein is generally down-regulated, was found.
Knutson
A
A
Department of Surgery, Uppsala University Hospital, Sweden.
Lillhager
P
P
Westin
G
G
D000236:Adenoma / Q000235:genetics*; D001483:Base Sequence; D018899:CpG Islands; D019175:DNA Methylation; D017931:DNA Primers; D026541:Heymann Nephritis Antigenic Complex; D006801:Humans; D008562:Membrane Glycoproteins / Q000235:genetics*; D010282:Parathyroid Neoplasms / Q000235:genetics*; D011401:Promoter Regions, Genetic; D014158:Transcription, Genetic / Q000235:genetics
D017931:DNA Primers; D026541:Heymann Nephritis Antigenic Complex; D008562:Membrane Glycoproteins