Roman-Roman S, Ferradini L, Azogui O, Faure F, Hercend T, Triebel F
Laboratoire d'Hémato-Immunologie, INSERM U333, Institut Gustave-Roussy, Villejuif, France.
Mol Immunol. 1993 Apr;30(5):423-31. doi: 10.1016/0161-5890(93)90110-w.
We used the anchored-polymerase chain reaction (A-PCR) procedure to study human TCR transcripts derived from a variety of polyclonal T cell populations. In this series of experiments, 31 'unusual' cDNAs, which do not include exclusively V-J-C, J-C or 5'C genomic sequences, were identified. Ten of these were found to represent distinct types of alternatively spliced TCR alpha transcripts whose structure is derived from unusual splicing of one, two or even three intervening intronic sequences. The splicing events led to either conservation of a novel exon in the mRNA structure (designated aE1 alpha-aE5 alpha) between the V-J and C segments or to deletion of the 3' V region-J segment. In three cases, the alternatively spliced exons (aE1 alpha-aE3 alpha) interrupt the open translational reading frame of the corresponding V-J alpha segment. Nineteen and two cDNA represent sterile C beta or C delta transcripts, respectively. Their structures are derived from the conservation of a non-translatable exon, aE1 beta or aE1 delta, which is precisely spliced at the 5' end of the corresponding C exon sequences. Interestingly, the 3' region of the aE1 beta sequence is homologous to the murine C beta 0 exon. Together, these results led to the characterization of nine novel exons in the TCR alpha, beta and delta loci.
我们采用锚定聚合酶链反应(A-PCR)方法来研究源自多种多克隆T细胞群体的人类TCR转录本。在这一系列实验中,鉴定出31个“异常”cDNA,它们并非仅包含V-J-C、J-C或5'C基因组序列。其中10个被发现代表不同类型的可变剪接TCRα转录本,其结构源自一个、两个甚至三个内含子序列的异常剪接。这些剪接事件导致mRNA结构中V-J和C区段之间保留一个新外显子(命名为aE1α-aE5α),或者导致3'V区-J区段缺失。在三个案例中,可变剪接外显子(aE1α-aE3α)中断了相应V-Jα区段的开放翻译阅读框。19个和2个cDNA分别代表无功能的Cβ或Cδ转录本。它们的结构源自一个不可翻译外显子aE1β或aE1δ的保留,该外显子在相应C外显子序列的5'端精确剪接。有趣的是,aE1β序列的3'区域与小鼠Cβ0外显子同源。总之,这些结果促成了TCRα、β和δ基因座中九个新外显子的表征。