T-cell receptor ζ mRNA with an alternatively spliced 3′ untranslated region is generated predominantly in the peripheral blood T cells of systemic lupus erythematosus patients
T-cell receptor ζ mRNA with an alternatively spliced 3′ untranslated region is generated predominantly in the peripheral blood T cells of systemic lupus erythematosus patients
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具有选择性剪接的 3 非翻译区的 T 细胞受体 z mRNA 主要在系统性红斑狼疮患者的外周血 T 细胞中产生
作者：
K. Tsuzaka;N. Onoda;K. Yoshimoto;Y. Setoyama;K. Suzuki;M. Pang;T. Abe;T. Takeuchi
摘要：为了探讨系统性红斑狼疮（SLE）患者外周血 T 细胞（PBT）中 T 细胞受体 z 链（TCR z）表达下调的机制，我们分析了 TCR z mRNA 的 3' 非翻译区（3'UTR），因为 mRNA 中的 3'UTR 负责转录后调节。使用逆转录酶聚合酶链式反应 (RT-PCR) 扩增 917bp TCRz 3′UTR cDNA 表明，短变异 cDNA (355bp)（表达为具有 562bp 缺失的选择性剪接 3′UTR）在 14 名 SLE 患者中的 11 名 PBT 中占主导地位，而野生型 cDNA (917bp) 则主要存在于 PBT 中。在七个阴性对照（两个系统性硬化症患者，五个正常对照）和两个 T 细胞系杂交瘤的 PBT 中检测到。半定量 PCR 还揭示了 SLE T 细胞中具有可变剪接 3'UTR 的 TCRz mRNA (TCRz mRNA/as-3'UTR) 的主要表达，以及具有野生形式 3'UTR 的 TCRz mRNA (TCRz mRNA/w-3'UTR) 的表达减少。然而，阴性对照和SLE患者之间开放阅读框（ORF）TCR z mRNA的表达没有差异。根据蛋白质印迹分析，TCR z 蛋白表达水平与 TCR z mRNA/w-3'UTR (r = 0.931) 良好相关，并且与 TCR z mRNA/as-3'UTR (r = -0.614) 可逆相关，但与 ORF TCR z mRNA 不相关 (r = -0.296)。由此可见，SLE T细胞中TCR z mRNA/w-3'UTR表达的减少和TCR z mRNA/as-3'UTR的显着表达导致TCR z蛋白的下调。
Abstract To investigate the mechanism of the downregulation of T-cell receptor ζ chain (TCRζ) expression in the peripheral blood T cells (PBTs) of systemic lupus erythematosus (SLE) patients, we analyzed the 3′ untranslated region (3′UTR) of TCRζ mRNA, because the 3′UTR in mRNA is responsible for posttranscriptional regulation. Use of the reverse transcriptase polymerase chain reaction (RT-PCR) to amplify the 917 bp TCRζ 3′UTR cDNA demonstrated that the short variant cDNA (355 bp), expressed as an alternatively spliced 3′UTR with 562-bp deletion, was predominated in the PBTs of 11 of 14 SLE patients, whereas mainly the wild-form cDNA (917 bp) was detected in the PBTs of seven negative controls (two systemic sclerosis patients, five normal controls) and in two T-cell line hybridomas. Semiquantitative PCR also revealed the predominant expression of the TCRζ mRNA with alternatively spliced 3′UTR (TCRζ mRNA/as-3′UTR), and a decreased expression of TCRζ mRNA with the wild form 3′UTR (TCRζ mRNA/w-3′UTR) in SLE T cells. However, there was no difference in the expression of the open reading frame (ORF) TCRζ mRNA between the negative controls and SLE patients. The TCRζ protein expression level according to Western blot analysis correlated well with that of TCRζ mRNA/w-3′UTR (r= 0.931) and reversibly with TCRζ mRNA/as-3′UTR (r=−0.614), but not with ORF TCRζ mRNA (r=−0.296). It can be concluded that the reduced expression of TCRζ mRNA/w-3′UTR and the predominant expression of TCRζ mRNA/as-3′UTR lead to downregulation of the TCRζ protein in SLE T cells.
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DOI：
10.1016/s0169-328x(98)00134-x
发表时间：
1998-08-15
期刊：
MOLECULAR BRAIN RESEARCH
影响因子：
--
作者：
Boado, RJ;Pardridge, WM
通讯作者：
Pardridge, WM
影响因子：
4.4
作者：
M. Berman;Christy Sandborg;Z. Wang;K. Imfeld;F. Zaldivar;V. Dadufalza;Bruce A. Buckingham
通讯作者：
M. Berman;Christy Sandborg;Z. Wang;K. Imfeld;F. Zaldivar;V. Dadufalza;Bruce A. Buckingham
DOI：
10.1073/pnas.95.25.14869
发表时间：
1998-12-08
影响因子：
11.1
作者：
Moser, KL;Neas, BR;Harley, JB
通讯作者：
Harley, JB
影响因子：
15.9
作者：
Tsao, BP;Cantor, RM;Notter, JI
通讯作者：
Notter, JI
DOI：
10.1073/pnas.95.25.14875
发表时间：
1998-12-08
影响因子：
11.1
作者：
Gaffney, PM;Kearns, GM;Behrens, TW
通讯作者：
Behrens, TW