The overall objective of the proposed research is to characterize the low density lipoprotein (LDL) receptor gene and to understand how it is altered in humans with familial hypercholesterolemia. Our initial goal will be the isolation of a complementary DNA clone to the bovine LDL receptor mRNA via the techniques of polyribosome immune precipitation and oligodeoxyreibonucleotide hybridization. We then propose to use this clone as a hybridization probe to identify human receptor cDNAs. Our second objective will be to characterize a full-length human cDNA clone at the primary structure level by standard DNA sequencing methodologies. From these results we will derive a series of oligodeoxyribonucleotides which span the entire cDNA and which are spaced approximaately 200 base pairs apart. In future studies, this series of primers will be used to define the lesions present in mutant receptor genes. Our third objective is the isolation and characterization of the human LDL receptor gene from a bacteriophage library. We will use the cDNA clones as probes to identify the gene in these recombinants and will characterize it with respect to size, restriction endonuclease mapping, and its exon-intron arrangement. The results of the above studies will provide a framework for interpreting changes which are present in mutant receptor genes isolated from familial hypercholesterolemic patients. The final objective is to characterize two different mutant LDL receptor genes that occur in these patients. These two mutations are of interest because they alter the size of the receptor protein and disrupt its processing and transport to the cell surfaces.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Modified Research Career Development Award (K04)
Project #
5K04HL001287-05
Application #
3073720
Study Section
Molecular Biology Study Section (MBY)
Project Start
1984-04-01
Project End
1989-03-31
Budget Start
1988-04-01
Budget End
1989-03-31
Support Year
5
Fiscal Year
1988
Total Cost
Indirect Cost
Name
University of Texas Sw Medical Center Dallas
Department
Type
Overall Medical
DUNS #
City
Dallas
State
TX
Country
United States
Zip Code
75390
Russell, D W; Esser, V; Hobbs, H H (1989) Molecular basis of familial hypercholesterolemia. Arteriosclerosis 9:I8-13
Andersson, S; Bishop, R W; Russell, D W (1989) Expression cloning and regulation of steroid 5 alpha-reductase, an enzyme essential for male sexual differentiation. J Biol Chem 264:16249-55
Cuthbert, J A; Russell, D W; Lipsky, P E (1989) Regulation of low density lipoprotein receptor gene expression in human lymphocytes. J Biol Chem 264:1298-304
Andersson, S; Davis, D L; Dahlback, H et al. (1989) Cloning, structure, and expression of the mitochondrial cytochrome P-450 sterol 26-hydroxylase, a bile acid biosynthetic enzyme. J Biol Chem 264:8222-9
Hofmann, S L; Russell, D W; Brown, M S et al. (1988) Overexpression of low density lipoprotein (LDL) receptor eliminates LDL from plasma in transgenic mice. Science 239:1277-81
Hobbs, H H; Leitersdorf, E; Goldstein, J L et al. (1988) Multiple crm- mutations in familial hypercholesterolemia. Evidence for 13 alleles, including four deletions. J Clin Invest 81:909-17
Esser, V; Russell, D W (1988) Transport-deficient mutations in the low density lipoprotein receptor. Alterations in the cysteine-rich and cysteine-poor regions of the protein block intracellular transport. J Biol Chem 263:13276-81
Russell, D W (1987) The study of natural and synthetic mutations in the LDL receptor. Kidney Int Suppl 23:S156-66
Sudhof, T C; Van der Westhuyzen, D R; Goldstein, J L et al. (1987) Three direct repeats and a TATA-like sequence are required for regulated expression of the human low density lipoprotein receptor gene. J Biol Chem 262:10773-9
Russell, D W (1987) Protein domains of the low density lipoprotein receptor. Acta Med Scand Suppl 715:39-44

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