caa a22 4500 397573847 CHVBK 20180308164849.0 cr unu---uuuuu 161202e199610 xx s 000 0 eng 10.1093/hmg/5.10.1581 doi (NATIONALLICENCE)oxford-10.1093/hmg/5.10.1581 Mutant Fibrillin-1 Monomers Lacking EGF-Like Domains Disrupt Microfibril Assembly and Cause Severe Marfan Syndrome [Elektronische Daten] [Wanguo Liu, Chiping Qian, Kimberly Comeau, Thomas Brenn, Heinz Furthmayr, Uta Francke] Marfan syndrome (MFS), a heritable connective tissue disorder, is caused by mutations in the gene coding for fibrillin-1 (FBN1), an extracellular matrix protein. One of the three major categories of FBN1 mutations involves exon-skipping. To rapidly detect such mutations, we developed a long RT-PCR method. Either three segments covering the entire FBN1 coding sequence or a single 8.9 kb FBN1 coding segment were amplified from reverse-transcribed total fibroblast RNA. Restriction fragment patterns of these RT-PCR products were compared and abnormal fragments were directly sequenced. Six exon-skipping mutations were identified in a panel of 60 MFS probands. All skipped exons encode calcium binding epidermal growth factor (EGF)-like domains and maintain the reading frame. In five probands, exon-skipping was due to point mutations in splice site sequences, and one had a 6 bp deletion in a donor splice site. Pulse-chase analysis of labelled fibrillin protein revealed normal levels of synthesis but significantly reduced matrix deposition. This dominant-negative effect of the mutant monomers is considered in the light of current models of fibrillin assembly. Probands with this type of FBN1 mutation include the most severe forms of MFS, such as neonatally lethal presentations. © 1996 Oxford University Press Liu Wanguo Howard Hughes Medical Institute, Stanford University Medical Center, Stanford, CA 94305, USA aut Qian Chiping Howard Hughes Medical Institute, Stanford University Medical Center, Stanford, CA 94305, USA aut Comeau Kimberly Howard Hughes Medical Institute, Stanford University Medical Center, Stanford, CA 94305, USA aut Brenn Thomas Department of Pathology, Stanford University Medical Center, Stanford, CA 94305, USA aut Furthmayr Heinz Department of Pathology, Stanford University Medical Center, Stanford, CA 94305, USA aut Francke Uta Howard Hughes Medical Institute, Stanford University Medical Center, Stanford, CA 94305, USA aut Human Molecular Genetics Oxford University Press 5/10(1996-10), 1581-1587 0964-6906 5:10<1581 1996 5 hmg https://doi.org/10.1093/hmg/5.10.1581 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1093/hmg/5.10.1581 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Liu Wanguo Howard Hughes Medical Institute, Stanford University Medical Center, Stanford, CA 94305, USA aut NATIONALLICENCE 700 1- Qian Chiping Howard Hughes Medical Institute, Stanford University Medical Center, Stanford, CA 94305, USA aut NATIONALLICENCE 700 1- Comeau Kimberly Howard Hughes Medical Institute, Stanford University Medical Center, Stanford, CA 94305, USA aut NATIONALLICENCE 700 1- Brenn Thomas Department of Pathology, Stanford University Medical Center, Stanford, CA 94305, USA aut NATIONALLICENCE 700 1- Furthmayr Heinz Department of Pathology, Stanford University Medical Center, Stanford, CA 94305, USA aut NATIONALLICENCE 700 1- Francke Uta Howard Hughes Medical Institute, Stanford University Medical Center, Stanford, CA 94305, USA aut NATIONALLICENCE 773 0- Human Molecular Genetics Oxford University Press 5/10(1996-10), 1581-1587 0964-6906 5:10<1581 1996 5 hmg Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford