Jolly C J, O'Neill H C
Molecular Immunology Laboratory, John Curtin School of Medical Research, Australian National University, Canberra.
Int Immunol. 1995 Jul;7(7):1147-56. doi: 10.1093/intimm/7.7.1147.
The TCR in a mature T cell is a multimeric complex of TCR alpha and beta chains, and CD3 subunits. Functional TCR alpha and beta chains are encoded by genes that result from developmentally controlled somatic rearrangement events. By FACS analysis, we have detected a TCR V beta 8 protein on the surface of an immature lymphoid cell line, C1-V13D, that has all of its TCR genes in germline (unrearranged) configuration. RNA blot analysis detected a 1.4 kb polydenylated V beta 8 RNA in C1-V13D cells, but no expression of C beta was detected. Rapid amplification of 3' cDNA ends was used to clone an RNA that was initiated from the leader exon of the V beta 5.1 gene and spliced to the V exon of the V beta 8.2 gene. The putative sequence of the mature 10.8 kDa protein was entirely encoded by the V beta 8.2 exon. RT-PCR analysis confirmed that 97% of the V beta 8 RNA detected in C1-V13D cells was encoded by the V beta 8.2 gene, and only 3% by V beta 8.1 and V beta 8.3 genes. Furthermore, most of the V beta 8.2 RNA was spliced to the leader exon of the V beta 5.1 gene and not to the leader exon of the V beta 8.2 gene. The implications of preferential transcription from particular germline TCR genes for repertoire diversity and possible functions for proteins translated from germline TCR V beta genes are discussed.
成熟T细胞中的TCR是由TCRα和β链以及CD3亚基组成的多聚体复合物。功能性TCRα和β链由发育控制的体细胞重排事件产生的基因编码。通过荧光激活细胞分选(FACS)分析,我们在未成熟淋巴细胞系C1-V13D的表面检测到一种TCR Vβ8蛋白,该细胞系所有的TCR基因都处于种系(未重排)构型。RNA印迹分析在C1-V13D细胞中检测到一个1.4 kb的多聚腺苷酸化Vβ8 RNA,但未检测到Cβ的表达。3' cDNA末端的快速扩增用于克隆一种RNA,该RNA从Vβ5.1基因的前导外显子起始并剪接到Vβ8.2基因的V外显子。推测的10.8 kDa成熟蛋白序列完全由Vβ8.2外显子编码。逆转录聚合酶链反应(RT-PCR)分析证实,在C1-V13D细胞中检测到的97%的Vβ8 RNA由Vβ8.2基因编码,只有3%由Vβ8.1和Vβ8.3基因编码。此外,大多数Vβ...