CADM1 is expressed as multiple alternatively spliced functional and dysfunctional isoforms in human mast cells.
CADM1 is expressed as multiple alternatively spliced functional and dysfunctional isoforms in human mast cells.
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DOI：
10.1016/j.molimm.2012.08.024
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发表时间：
2013-04
影响因子：
3.6
通讯作者：
Bradding P
中科院分区：
文献类型：
--
作者：
Moiseeva EP;Leyland ML;Bradding P
人肥大细胞表达6种选择性剪接的CADM 1亚型：4种功能性和2种功能障碍性。CADM 1 SP1和SP 6仅限于分化的人肺肥大细胞。混合同种型的表达减少人肥大细胞与肺成纤维细胞的粘附。在40%的肥大细胞中发现了CADM 1亚型的功能障碍。CADM 1功能障碍的亚型限制CADM 1的表达，并可能导致各种疾病。细胞粘附分子1（CADM 1）参与多种疾病的发病机制，并负责肥大细胞（MC）的粘附和存活。CADM 1亚型在不同物种中的表达存在差异。我们以前克隆了SP 4，SP1，SP 6和一个功能障碍的亚型从人肺MC（HLMC）和MC系HMC-1。本研究的目的是鉴定人MC中表达的所有亚型。RT-PCR检测到人MCs中存在SP 4、SP1、SP 6和SP3的功能亚型，这些亚型在外显子7/11之间存在选择性剪接。两个功能失调的异构体与隐性外显子A和B之间的外显子1/2的选择性剪接，导致翻译的提前终止，发现在约40%的MC标本。基因组DNA测序结果表明，隐藏外显子B的剪接不是由该外显子或其假定剪接分支点内的特异性SNP引起的。蛋白质印迹法检测到高度糖基化的CADM 1（约105 kDa），但仅在HLMCs的培养基中发现胞外结构域（约95 kDa），而在HMC-1细胞中未发现，表明差异蛋白质表达。转染SP1和SP 6，而不是SP 4，减少HMC-1细胞粘附到人肺成纤维细胞，但不是气道平滑肌细胞。因此，在人类MC中发现了功能障碍性和功能性CADM 1同种型。较长的SP1和SP 6在分化的HLMCs中最明显，与SP 4相比显示出差异性粘附。这些多种异构体可能有助于健康和疾病中的MC功能。
► Human mast cells express 6 alternatively spliced CADM1 isoforms: 4 functional and 2 dysfunctional. ► Long CADM1 SP1 and SP6 are restricted to differentiated human lung mast cells. ► Expression of mixed isoforms reduces adhesion of human mast cells to lung fibroblasts. ► Dysfunctional CADM1 isoforms are found in 40% of mast cells. ► Dysfunctional isoforms limit CADM1 expression and may predispose to various diseases. Cell adhesion molecule 1 (CADM1) is implicated in the pathogenesis of several diseases and is responsible for adhesion and survival of mast cells (MCs). Differential expression of CADM1 isoforms was found in different species. We previously cloned SP4, SP1, SP6 and a dysfunctional isoform from human lung MCs (HLMCs) and the MC line HMC-1. The aim of this study was to identify all isoforms expressed in human MCs. The functional isoforms SP4, SP1, SP6 and SP3, with alternative splicing between exons 7/11, were detected in human MCs by RT-PCR. Two dysfunctional isoforms with alternative splicing of cryptic exons A and B between exons 1/2, leading to premature termination of translation, were found in ∼40% of MC specimens. Sequencing of genomic DNA showed that splicing of cryptic exon B did not result from specific SNPs within this exon or its putative splice branch point. Highly glycosylated CADM1 (∼105 kDa) was detected by western blotting, but an extracellular domain (∼95 kDa) was found only in the culture medium from HLMCs, but not HMC-1 cells, indicating differential protein expression. Transfection of SP1 and SP6, but not SP4, reduced adhesion of HMC-1 cells to human lung fibroblasts but not airway smooth muscle cells. Hence, dysfunctional and functional CADM1 isoforms are found in human MCs. The longer SP1 and SP6 were most evident in differentiated HLMCs and displayed differential adhesion compared to SP4. These multiple isoforms are likely to contribute to MC function in both health and disease.
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DOI：
10.4049/jimmunol.181.4.2772
发表时间：
2008-08-15
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Hollins F;Kaur D;Yang W;Cruse G;Saunders R;Sutcliffe A;Berger P;Ito A;Brightling CE;Bradding P
通讯作者：
Bradding P
影响因子：
5.4
作者：
Dewan, M. Zahidunnabi;Takamatsu, Naofumi;Morishita, Kazuhiro
通讯作者：
Morishita, Kazuhiro
DOI：
10.1164/rccm.200603-394oc
发表时间：
2006-12-01
影响因子：
24.7
作者：
Kaur, Davinder;Saunders, Ruth;Brightling, Christopher E.
通讯作者：
Brightling, Christopher E.
影响因子：
4.8
作者：
Cruse, Glenn;Kaur, Davinder;Bradding, Peter
通讯作者：
Bradding, Peter
影响因子：
20.3
作者：
Hasstedt, Sandra J.;Bezemer, Irene D.;Bovill, Edwin G.
通讯作者：
Bovill, Edwin G.