دورية أكاديمية
أعرض في EDS

Mechanical Stress and Single Nucleotide Variants Regulate Alternative Splicing of the MYLK Gene.

التفاصيل البيبلوغرافية
العنوان: Mechanical Stress and Single Nucleotide Variants Regulate Alternative Splicing of the MYLK Gene.
المؤلفون: Mascarenhas JB; 1 Department of Medicine, and., Tchourbanov AY; 2 Arizona Research Laboratory, University of Arizona, Tucson, Arizona; and., Fan H; 3 Department of Anesthesiology, University of Illinois at Chicago, Chicago, Illinois., Danilov SM; 1 Department of Medicine, and.; 3 Department of Anesthesiology, University of Illinois at Chicago, Chicago, Illinois., Wang T; 1 Department of Medicine, and., Garcia JG; 1 Department of Medicine, and.
المصدر: American journal of respiratory cell and molecular biology [Am J Respir Cell Mol Biol] 2017 Jan; Vol. 56 (1), pp. 29-37.
نوع المنشور: Journal Article
اللغة: English
بيانات الدورية: Publisher: American Thoracic Society Country of Publication: United States NLM ID: 8917225 Publication Model: Print Cited Medium: Internet ISSN: 1535-4989 (Electronic) Linking ISSN: 10441549 NLM ISO Abbreviation: Am J Respir Cell Mol Biol Subsets: MEDLINE
أسماء مطبوعة: Publication: 2000- : New York, NY : American Thoracic Society
Original Publication: [New York, NY : The Association, [c1989-
مواضيع طبية MeSH: Stress, Mechanical*, Alternative Splicing/*genetics , Myosin-Light-Chain Kinase/*genetics , Polymorphism, Single Nucleotide/*genetics, Base Sequence ; Computational Biology ; Computer Simulation ; Consensus Sequence/genetics ; Exons/genetics ; HEK293 Cells ; Humans ; Lymphocytes/metabolism ; RNA Splice Sites/genetics ; Reproducibility of Results
مستخلص: The nonmuscle (nm) myosin light-chain kinase isoform (MLCK), encoded by the MYLK gene, is a vital participant in regulating vascular barrier responses to mechanical and inflammatory stimuli. We determined that MYLK is alternatively spliced, yielding functionally distinct nmMLCK splice variants including nmMLCK2, a splice variant highly expressed in vascular endothelial cells (EC) and associated with reduced EC barrier integrity. We demonstrated previously that the nmMLCK2 variant lacks exon 11, which encodes a key regulatory region containing two differentially phosphorylated tyrosine residues (Y 464 and Y 471 ) that influence vascular barrier function during inflammation. In this study, we used minigene constructs and RT-PCR to interrogate biophysical factors (mechanical stress) and genetic variants (MYLK single-nucleotide polymorphisms [SNPs]) that are potentially involved in regulating MYLK alternative splicing and nmMLCK2 generation. Human lung EC exposed to pathologic mechanical stress (18% cyclic stretch) produced increased nmMLCK2 expression relative to levels of nmMLCK1 with alternative splicing significantly influenced by MYLK SNPs rs77323602 and rs147245669. In silico analyses predicted that these variants would alter exon 11 donor and acceptor sites for alternative splicing, computational predictions that were confirmed by minigene studies. The introduction of rs77323602 favored wild-type nmMLCK expression, whereas rs147245669 favored alternative splicing and deletion of exon 11, yielding increased nmMLCK2 expression. Finally, lymphoblastoid cell lines selectively harboring these MYLK SNPs (rs77323602 and rs147245669) directly validated SNP-specific effects on MYLK alternative splicing and nmMLCK2 generation. Together, these studies demonstrate that mechanical stress and MYLK SNPs regulate MYLK alternative splicing and generation of a splice variant, nmMLCK2, that contributes to the severity of inflammatory injury.
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معلومات مُعتمدة: P01 HL058064 United States HL NHLBI NIH HHS; P01 HL126609 United States HL NHLBI NIH HHS; R01 HL091889 United States HL NHLBI NIH HHS
فهرسة مساهمة: Keywords: bioinformatics; cyclic stretch; single nucleotide polymorphism; splicing; ventilator-induced lung injury
المشرفين على المادة: 0 (RNA Splice Sites)
EC 2.7.11.18 (Myosin-Light-Chain Kinase)
تواريخ الأحداث: Date Created: 20160817 Date Completed: 20170630 Latest Revision: 20181113
رمز التحديث: 20231215
مُعرف محوري في PubMed: PMC5248959
DOI: 10.1165/rcmb.2016-0053OC
PMID: 27529643
قاعدة البيانات: MEDLINE
الوصف
تدمد:1535-4989
DOI:10.1165/rcmb.2016-0053OC