Detection of spliced and unspliced forms of germline TCR-Vbeta transcripts in extrathymic lymphoid sites.
Germline TCR-Vbeta transcription is commonly considered an event coupled with rearrangement of TCR genes in T cells. The extent of germline Vbeta transcription is studied here in a range of cell types and in several mouse strains. A sensitive semi-quantitative RT-PCR method was developed to specifically detect germline and not rearranged transcripts. Germline transcription of a range of different Vbeta genes was detected along with rearranged transcripts in bone marrow, thymus, mesenteric lymph node and spleen. Some transcripts were also detected in low level in non-lymphoid tissues including heart, liver and brain. Expression was also studied in the C57BL/6J-beta2microglobulin-/- (C57BL/6J-beta2M-/-) mouse model that lacks NK1.1 T cells and predominantly utilises Vbeta8.2 in the formation of a TCR. beta2M-/- mice, which lack both CD1-dependent NK1.1 T cells and CD8+ T cells, showed germline TCR-Vbeta8 transcription in most tissues indicating that germline transcription is not specifically related to CD1-dependent NK1.1 T cells. In many tissues, multiple transcripts were amplified representing both spliced and unspliced forms of germline Vbeta. For most Vbeta genes, the expression of spliced and unspliced forms was equivalent. Given an abundance of unspliced transcripts, the presence of alternative ORFs encoding a novel protein was investigated within the TCR-Vbeta genes. Sequence analysis of ORFs showed only genes with a high level of similarity to TCR-beta. All data reflect the prevalence of germline transcripts in vivo and raise questions about their functional role.
Abbey
Janice L
JL
School of Biochemistry & Molecular Biology, Australian National University, Canberra, ACT 0200, Australia.
O'Neill
Helen C
HC
D000818:Animals; D001483:Base Sequence; D030541:Databases, Genetic; D015333:Gene Rearrangement, beta-Chain T-Cell Antigen Receptor; D007438:Introns; D008221:Lymphoid Tissue / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D008817:Mice, Mutant Strains; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D009928:Organ Specificity; D012326:RNA Splicing; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013045:Species Specificity
D016693:Receptors, Antigen, T-Cell, alpha-beta