Adaptor protein disabled-2 modulates low density lipoprotein receptor synthesis in fibroblasts from patients with autosomal recessive hypercholesterolaemia

Hum Mol Genet. 2007 Nov 15;16(22):2751-9. doi: 10.1093/hmg/ddm232. Epub 2007 Aug 29.

Abstract

Autosomal recessive hypercholesterolaemia (ARH), characterized clinically by severe inherited hypercholesterolaemia, is caused by recessive null mutations in LDLRAP1 (formerly ARH). Immortalized lymphocytes and monocyte-macrophages, and presumably hepatocytes, from ARH patients fail to take up and degrade plasma low density lipoproteins (LDL) because they lack LDLRAP1, a cargo-specific adaptor required for clathrin-mediated endocytosis of the LDL receptor. Surprisingly, LDL-receptor function is normal in ARH patients' skin fibroblasts in culture. Disabled-2 (Dab2) has been implicated previously in clathrin-mediated internalization of LDL-receptor family members, and we show here that Dab2 is highly expressed in skin fibroblasts, but not in lymphocytes. SiRNA-depletion of Dab2 profoundly reduced LDL-receptor activity in ARH fibroblasts as a result of profound reduction in LDL-receptor protein, but not mRNA; heterologous expression of murine Dab2 reversed this effect. In contrast, LDL-receptor protein content was unchanged in Dab-2-depleted control cells. Incorporation of 35S-labelled amino acids into LDL receptor protein revealed a corresponding apparent reduction in accumulation of newly synthesized LDL-receptor protein on depletion of Dab2 in ARH, but not in control, cells. This reduction in LDL-receptor protein in Dab2-depleted ARH cells could not be reversed by treatment of the cells with proteasomal or lysosomal inhibitors. Thus, we propose a novel role for Dab2 in ARH fibroblasts, where it is apparently required to allow normal translation of LDL receptor mRNA.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Protein Complex 2 / genetics
  • Adaptor Protein Complex 2 / metabolism*
  • Adaptor Proteins, Signal Transducing / genetics*
  • Adaptor Proteins, Signal Transducing / metabolism
  • Animals
  • CHO Cells
  • Case-Control Studies
  • Cricetinae
  • Cricetulus
  • Endocytosis
  • Fibroblasts / metabolism*
  • Fibroblasts / pathology
  • Genes, Recessive
  • Humans
  • Hyperlipoproteinemia Type II / genetics*
  • Lymphocytes / metabolism
  • RNA, Messenger / genetics
  • RNA, Small Interfering
  • Receptors, LDL / genetics
  • Receptors, LDL / metabolism*
  • Skin / metabolism*
  • Skin / pathology
  • Transcription, Genetic

Substances

  • Adaptor Protein Complex 2
  • Adaptor Proteins, Signal Transducing
  • LDLRAP1 protein, human
  • RNA, Messenger
  • RNA, Small Interfering
  • Receptors, LDL