The enzyme HMG CoA reductase catalyzes conversion of HMG CoA to mevalonate, a rate-limiting step in synthesis of cholesterol and essential nonsterol isoprenoids. Reductase is anchored to endoplasmic reticulum (ER) membranes through an N-terminal domain that contains eight membrane-spanning helices;the C- terminal domain projects into the cytosol and exerts catalytic activity. Accelerated ER-associated degradation (ERAD), one of several mechanisms for feedback control of reductase, results from sterol-induced binding of reductase to ER membrane proteins called Insig-1 and Insig-2. Ubiquitination of reductase by Insig-associated ubiquitin ligases marks the enzyme for retrotranslocation across ER membranes and dislocation into the cytosol for proteasomal degradation. These postubiquitination steps are modulated by the nonsterol isoprenoid geranylgeraniol through a completely unknown mechanism. In preliminary studies, we find that sterols also trigger binding of reductase to UbiA prenyltransferase domain containing protein-1 (UBIAD1), which catalyzes geranylgeranylation of bacterial and plant vitamin K derivatives to produce menaquinone-4 (MK-4). Mutations in UBIAD1 cause a rare autosomal dominant eye disease called Schnyder corneal dystrophy (SCD), which is characterized by progressive accumulation of cholesterol in the cornea. Sterol-induced binding of reductase to UBIAD1 is inhibited by geranylgeraniol and RNA interference studies reveal that reductase-UBIAD1 binding leads to Insig recruitment and ERAD of reductase. Thus, we hypothesize that 1) UBIAD1 mediates the sterol-sensing reaction required for reductase-Insig binding;and 2) geranylgeraniol-mediated displacement of UBIAD1 is required for initiation of postubiquitination steps in reductase ERAD. To appraise these hypotheses, we propose three Specific Aims: 1) delineate mechanisms by which UBIAD1 mediates sterol-accelerated ERAD of reductase;2) explore mechanism through which sterols modulate function of UBIAD1;and 3) determine the role of UBIAD1 in ERAD of reductase and regulation of cholesterol homeostasis in whole animals. Collectively, these studies will provide key information regarding the removal of polytopic proteins from ER membranes for proteasomal degradation. In addition, these studies have significant clinical implications. Reductase is the target of statins, widely prescribed drugs that lower plasma LDL-cholesterol and reduce the incidence of cardiovascular disease. Statins trigger responses that cause accumulation of reductase, which blunts their clinical effects. Part of this increase results from slowed ERAD of reductase. Thus, elucidating mechanisms for reductase ERAD holds promise for development of new therapies that increase the effectiveness of statins and ultimately reduce the incidence of heart attacks. Moreover, insight into mechanisms for reductase ERAD may lead to therapeutic interventions that retard or prevent corneal accumulation of cholesterol associated with SCD.

Public Health Relevance

Cholesterol is an essential component of cell membranes;its synthesis requires the action of more than 20 enzymes. The key enzyme is HMG CoA reductase, which is tightly controlled through multiple mechanisms, including regulation of protein stability. Statins, competitive inhibitors of HMG CoA reductase, are routinely used to lower blood cholesterol in humans. However, these drugs trigger responses that result in accumulation of reductase protein, which often blunts their cholesterol-lowering effects. Part of this increase in reductase is due to slowed degradation of the enzyme. This grant will investigate mechanisms for reductase degradation, the elucidation of which will provide insight into development of new therapies that counteract statin-induced accumulation of reductase and improve the effectiveness of the drugs. Moreover, these studies will provide insight into therapies to prevent the corneal accumulation of cholesterol associated with the eye disease Schnyder corneal dystrophy.

Agency
National Institute of Health (NIH)
Type
Research Project (R01)
Project #
1R01GM112409-01
Application #
8786245
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Gindhart, Joseph G
Project Start
Project End
Budget Start
Budget End
Support Year
1
Fiscal Year
2014
Total Cost
Indirect Cost
Name
University of Texas Sw Medical Center Dallas
Department
Genetics
Type
Schools of Medicine
DUNS #
City
Dallas
State
TX
Country
United States
Zip Code
75390