HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Lelli, N. Articles by Calandra, S. Search for Related Content PubMed PubMed Citation Articles by Lelli, N. Articles by Calandra, S. Social Bookmarking                      What's this?

Journal of Lipid Research, Vol 36, 1315-1324, Copyright © 1995 by Lipid Research, Inc.

ARTICLES

Occurrence of multiple aberrantly spliced mRNAs of the LDL-receptor gene upon a donor splice site mutation that causes familial hypercholesterolemia (FHBenevento)

N Lelli, R Garuti, M Ghisellini, R Tiozzo, M Rolleri, V Aimale, E Ginocchio, A Naselli, S Bertolini and S Calandra
Dipartimento di Scienze Biomediche, Universita di Modena, Italy.

A novel point mutation of the LDL-receptor gene was found in an Italian patient with homozygous familial hypercholesterolemia. The SSCP analysis of the promoter and of 16 out of the 18 exons of the LDL- receptor gene was negative, suggesting that the mutation might be located in the region of the gene encompassing exons 14 and 15, a region that had not been amenable to polymerase chain reaction (PCR) amplification from genomic DNA. This region was amplified from cDNA by reverse transcription PCR (RT-PCR). RT-PCR of proband cDNA generated three fragments of 800, 600, and 550 bp, respectively, as opposed to a single 720 bp fragment obtained from control cDNA. The sequence of these fragments showed that: i) in the 800-bp fragment exon 14 continued with the 5' end of intron 15 (90 nucleotides), which in turn was followed by exon 16; ii) in the 600-bp fragment exon 14 was followed by the 5' end of exon 15 (50 nucleotides), which continued with exon 16; iii) in the 550-bp fragment exon 14 joined directly to exon 16. These abnormally spliced mRNAs resulted from a G-->A transition at the +1 nucleotide of intron 15, which changed the invariant GT dinucleotide of the 5' donor splice site. That was associated with the activation of two cryptic donor splice sites in intron 15 and exon 15, respectively, and the use of an alternative splicing leading to the skipping of exon 15. Northern blot analysis showed that the overall content of these aberrantly spliced mRNAs in proband fibroblasts was one-fourth that found in control cells. These abnormally spliced mRNAs are predicted to encode three abnormal receptor proteins: the first would contain an insertion of 30 novel amino acids; the second would be a truncated protein of 709 amino acids; the third would be devoid of the 57 amino acids of the O-linked sugar domain. Ligand blot experiments indicated that the amount of LDL- receptor present in proband's fibroblasts was approximately one-tenth that found in control cells.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				S. Bertolini, A. Cantafora, M. Averna, C. Cortese, C. Motti, S. Martini, G. Pes, A. Postiglione, C. Stefanutti, I. Blotta, et al. Clinical Expression of Familial Hypercholesterolemia in Clusters of Mutations of the LDL Receptor Gene That Cause a Receptor-Defective or Receptor-Negative Phenotype Arterioscler. Thromb. Vasc. Biol., September 1, 2000; 20 (9): e41 - e52. [Abstract] [Full Text] [PDF]

				Y. von Kodolitsch, R. E. Pyeritz, and P. K. Rogan Splice-Site Mutations in Atherosclerosis Candidate Genes : Relating Individual Information to Phenotype Circulation, August 17, 1999; 100(7): 693 - 699. [Abstract] [Full Text] [PDF]

				S. Bertolini, S. Cassanelli, R. Garuti, M. Ghisellini, M. L. Simone, M. Rolleri, P. Masturzo, and S. Calandra Analysis of LDL Receptor Gene Mutations in Italian Patients With Homozygous Familial Hypercholesterolemia Arterioscler. Thromb. Vasc. Biol., February 1, 1999; 19(2): 408 - 418. [Abstract] [Full Text] [PDF]

				Dilip. D. Patel, N. Lelli, R. Garuti, S. L. Volti, S. Bertolini, B. L. Knight, and S. Calandra Analysis of two duplications of the LDL receptor gene affecting intracellular transport, catabolism, and surface binding of the LDL receptor J. Lipid Res., July 1, 1998; 39(7): 1466 - 1475. [Abstract] [Full Text]

				T. Sampietro, M. Tuoni, M. Ferdeghini, A. Ciardi, P. Marraccini, C. Prontera, G. Sassi, M. Taddei, and A. Bionda Plasma Cholesterol Regulates Soluble Cell Adhesion Molecule Expression in Familial Hypercholesterolemia Circulation, September 2, 1997; 96(5): 1381 - 1385. [Abstract] [Full Text]