دورية أكاديمية
Functional Characterization of a Novel Truncating Mutation in Lamin A/C Gene in a Family with a Severe Cardiomyopathy with Conduction Defects
| العنوان: | Functional Characterization of a Novel Truncating Mutation in Lamin A/C Gene in a Family with a Severe Cardiomyopathy with Conduction Defects |
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| المؤلفون: | Andrea Gerbino, Irene Bottillo, Serena Milano, Martina Lipari, Roberta De Zio, Silvia Morlino, Maria Grazia Mola, Giuseppe Procino, Federica Re, Elisabetta Zachara, Paola Grammatico, Maria Svelto, Monica Carmosino |
| المصدر: | Cellular Physiology and Biochemistry, Vol 44, Iss 4, Pp 1559-1577 (2017) |
| بيانات النشر: | Cell Physiol Biochem Press GmbH & Co KG, 2017. |
| سنة النشر: | 2017 |
| المجموعة: | LCC:Physiology LCC:Biochemistry |
| مصطلحات موضوعية: | Laminopathies, Nucleus, Endoplasmic reticulum, Connexin, Ca2+ signaling, Cardiomyocytes, Lamin A/C gene, Physiology, QP1-981, Biochemistry, QD415-436 |
| الوصف: | Background/Aims: Truncating LMNA gene mutations occur in many inherited cardiomyopathy cases, but the molecular mechanisms involved in the disease they cause have not yet been systematically investigated. Here, we studied a novel frameshift LMNA variant (p.D243Gfs*4) identified in three members of an Italian family co-segregating with a severe form of cardiomyopathy with conduction defects. Methods: HEK293 cells and HL-1 cardiomyocytes were transiently transfected with either Lamin A or D243Gfs*4 tagged with GFP (or mCherry). D243Gfs*4 expression, cellular localization and its effects on diverse cellular mechanisms were evaluated with western blotting, laser-scanning confocal microscopy and video-imaging analysis in single cells. Results: When expressed in HEK293 cells, GFP- (or mCherry)-tagged LMNA D243Gfs*4 colocalized with calnexin within the ER. ER mislocalization of LMNA D243Gfs*4 did not significantly induce ER stress response, abnormal Ca2+ handling and apoptosis when compared with HEK293 cells expressing another truncated mutant of LMNA (R321X) which similarly accumulates within the ER. Of note, HEK293-LMNA D243Gfs*4 cells showed a significant reduction of connexin 43 (CX43) expression level, which was completely rescued by activation of the WNT/β-catenin signaling pathway. When expressed in HL-1 cardiomyocytes, D243Gfs*4 significantly impaired the spontaneous Ca2+ oscillations recorded in these cells as result of propagation of the depolarizing waves through the gap junctions between non-transfected cells surrounding a cell harboring the mutation. Furthermore, mCh-D243Gfs*4 HL-1 cardiomyocytes showed reduced CX43-dependent Lucifer Yellow (LY) loading and propagation. Of note, activation of β-catenin rescued both LY loading and LMNA D243Gfs*4 -HL-1 cells spontaneous activity propagation. Conclusion: Overall, the present results clearly indicate the involvement of the aberrant CX43 expression/activity as a pathogenic mechanism for the conduction defects associated to this LMNA truncating alteration. |
| نوع الوثيقة: | article |
| وصف الملف: | electronic resource |
| اللغة: | English |
| تدمد: | 1015-8987 1421-9778 |
| العلاقة: | https://www.karger.com/Article/FullText/485651Test; https://doaj.org/toc/1015-8987Test; https://doaj.org/toc/1421-9778Test |
| DOI: | 10.1159/000485651 |
| الوصول الحر: | https://doaj.org/article/0303a5d654164150b0089dedd1ad0c82Test |
| رقم الانضمام: | edsdoj.0303a5d654164150b0089dedd1ad0c82 |
| قاعدة البيانات: | Directory of Open Access Journals |
| تدمد: | 10158987 14219778 |
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| DOI: | 10.1159/000485651 |