Expression of a TCR V beta 8.2 polypeptide from the unrearranged gene in a murine lymphoid precursor cell line.
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.
Molecular Immunology Laboratory, John Curtin School of Medical Research, Australian National University, Canberra.
C J
CJ
Jolly
H C
HC
O'Neill
D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D015329:Gene Rearrangement, T-Lymphocyte; D051379:Mice; D008806:Mice, Inbred AKR; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D008969:Molecular Sequence Data; D010455:Peptides / Q000235:genetics* / Q000276:immunology; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000302:isolation & purification; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000235:genetics* / Q000276:immunology; D013194:Staining and Labeling; D013234:Stem Cells / Q000276:immunology*; D014158:Transcription, Genetic
D000911:Antibodies, Monoclonal; D010455:Peptides; D012333:RNA, Messenger; D016693:Receptors, Antigen, T-Cell, alpha-beta