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Blood, 1 November 2004, Vol. 104, No. 9, pp. 2739-2745. Prepublished online as a Blood First Edition Paper on June 29, 2004; DOI 10.1182/blood-2003-12-4286. This Article Full Text (PDF) All Versions of this Article: 2003-12-4286v1 104/9/2739    most recent Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Rights and Permissions Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by James, P. D Articles by Lillicrap, D. Search for Related Content PubMed PubMed Citation Articles by James, P. D Articles by Lillicrap, D. Social Bookmarking                   What's this?  Previous Article  |  Next Article  Submitted January 8, 2004 Accepted June 2, 2004 A Novel Type 2A von Willebrand Factor Mutation Located at the Last Nucleotide of Exon 26 (3538G>A) Causes Skipping of Two Non-Adjacent Exons Paula D James, Lee A O'Brien, Carol A Hegadorn, Colleen R Notley, Gary D Sinclair, Christine Hough, Man-Chiu Poon, and David Lillicrap* Pathology and Molecular Medicine, Queen's University, Kingston, Ontario, Canada Biochemistry and Molecular Biology, University of Calgary, Calgary, Alberta, Canada Medicine, Pediatrics, and Oncology, University of Calgary, Calgary, Alberta, Canada * Corresponding author; email: lillicrap{at}cliff.path.queensu.ca. In this manuscript, we describe a case of Type 2A VWD caused by the novel heterozygous GA transition at nucleotide 3538, which should result in the putative, non-conservative substitution of G1180R. This mutation was reproduced by site-directed mutagenesis; however, the recombinant mutant protein was efficiently secreted from cells and assembled correctly into multimers. Because the substitution is located at the last nucleotide of exon 26, the patient's platelet VWF mRNA was analyzed and three transcripts were observed: the normal transcript without the 3538G>A transition, a transcript with the inframe deletion of exon 26 and a transcript with the inframe deletions of exons 23 and 26. These deletion VWF cDNA constructs were created and the resulting recombinant proteins were analyzed following transfection into COS-7 cells. Co-transfection results demonstrate that the exon skipped transcripts lead to intracellular retention and the levels of VWF:Ag produced by these constructs were as follows: del23/26 < del26 < G1180R < /= wild type. The homozygous exon skipped transcripts show the presence of only the lowest molecular weight multimers. The G>A transition at nt 3538 does not result in the expression of the G1180R missense mutation, but rather leads to exon skipping which is the pathogenic basis of the patient's phenotype. This is the first report of a coding region mutation resulting in the skipping of 2 non-adjacent exons. CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this? This article has been cited by other articles: J. Wang, P. J. Smith, A. R. Krainer, and M. Q. Zhang Distribution of SR protein exonic splicing enhancer motifs in human protein-coding genes Nucleic Acids Res., September 7, 2005; 33(16): 5053 - 5062. [Abstract] [Full Text] [PDF] O. G. Ottmann and B. Wassmann Treatment of Philadelphia Chromosome-Positive Acute Lymphoblastic Leukemia Hematology, January 1, 2005; 2005(1): 118 - 122. [Abstract] [Full Text] [PDF]

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