Expression of a TCR Vβ8.2 polypeptide from the unrearranged gene in a murine lymphoid precursor cell line

Christopher J. Jolly, Helen C. O'neill

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10 Citations (Scopus)

Abstract

The TCR in a mature T cell is a multimeric complex of TCR α and β chains, and CD3 subunits. Functional TCR α and β chains are encoded by genes that result from developmentally controlled somatic rearrangement events. By FACS analysis, we have detected a TCR Vβ8 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 Vβ8 RNA in C1-V13D cells, but no expression of Cβ was detected. Rapid amplification of 3' cDNA ends was used to clone an RNA that was initiated from the leader exon of the Vβ5.1 gene and spliced to the V exon of the Vβ8.2 gene. The putative sequence of the mature 10.8 kDa protein was entirely encoded by the Vβ8.2 exon. RT-PCR analysis confirmed that 97% of the Vβ8 RNA detected in C1-V13D cells was encoded by the Vβ8.2 gene, and only 3% by Vβ8.1 and Vβ8.3 genes. Furthermore, most of the Vβ8.2 RNA was spliced to the leader exon of the Vβ5.1 gene and not to the leader exon of the Vβ8.2gene. The implications of preferential transcription from particular germline TCR genes for repertoire diversity and possible functions for proteins translated from germline TCR Vβ genes are discussed.

Original languageEnglish
Pages (from-to)1147-1156
Number of pages10
JournalInternational Immunology
Volume7
Issue number7
DOIs
Publication statusPublished - Jul 1995
Externally publishedYes

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Lymphocytes
Cell Line
Peptides
Exons
Genes
RNA
Spliced Leader RNA
T-Cell Antigen Receptor-CD3 Complex
Recombinant DNA
Membrane Proteins
Proteins
Complementary DNA
Clone Cells
T-Lymphocytes
Polymerase Chain Reaction

Cite this

@article{33ae3df51eec4aa28ebab9da29175f53,
title = "Expression of a TCR Vβ8.2 polypeptide from the unrearranged gene in a murine lymphoid precursor cell line",
abstract = "The TCR in a mature T cell is a multimeric complex of TCR α and β chains, and CD3 subunits. Functional TCR α and β chains are encoded by genes that result from developmentally controlled somatic rearrangement events. By FACS analysis, we have detected a TCR Vβ8 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 Vβ8 RNA in C1-V13D cells, but no expression of Cβ was detected. Rapid amplification of 3' cDNA ends was used to clone an RNA that was initiated from the leader exon of the Vβ5.1 gene and spliced to the V exon of the Vβ8.2 gene. The putative sequence of the mature 10.8 kDa protein was entirely encoded by the Vβ8.2 exon. RT-PCR analysis confirmed that 97{\%} of the Vβ8 RNA detected in C1-V13D cells was encoded by the Vβ8.2 gene, and only 3{\%} by Vβ8.1 and Vβ8.3 genes. Furthermore, most of the Vβ8.2 RNA was spliced to the leader exon of the Vβ5.1 gene and not to the leader exon of the Vβ8.2gene. The implications of preferential transcription from particular germline TCR genes for repertoire diversity and possible functions for proteins translated from germline TCR Vβ genes are discussed.",
author = "Jolly, {Christopher J.} and O'neill, {Helen C.}",
year = "1995",
month = "7",
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language = "English",
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Expression of a TCR Vβ8.2 polypeptide from the unrearranged gene in a murine lymphoid precursor cell line. / Jolly, Christopher J.; O'neill, Helen C.

In: International Immunology, Vol. 7, No. 7, 07.1995, p. 1147-1156.

Research output: Contribution to journalArticleResearchpeer-review

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T1 - Expression of a TCR Vβ8.2 polypeptide from the unrearranged gene in a murine lymphoid precursor cell line

AU - Jolly, Christopher J.

AU - O'neill, Helen C.

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N2 - The TCR in a mature T cell is a multimeric complex of TCR α and β chains, and CD3 subunits. Functional TCR α and β chains are encoded by genes that result from developmentally controlled somatic rearrangement events. By FACS analysis, we have detected a TCR Vβ8 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 Vβ8 RNA in C1-V13D cells, but no expression of Cβ was detected. Rapid amplification of 3' cDNA ends was used to clone an RNA that was initiated from the leader exon of the Vβ5.1 gene and spliced to the V exon of the Vβ8.2 gene. The putative sequence of the mature 10.8 kDa protein was entirely encoded by the Vβ8.2 exon. RT-PCR analysis confirmed that 97% of the Vβ8 RNA detected in C1-V13D cells was encoded by the Vβ8.2 gene, and only 3% by Vβ8.1 and Vβ8.3 genes. Furthermore, most of the Vβ8.2 RNA was spliced to the leader exon of the Vβ5.1 gene and not to the leader exon of the Vβ8.2gene. The implications of preferential transcription from particular germline TCR genes for repertoire diversity and possible functions for proteins translated from germline TCR Vβ genes are discussed.

AB - The TCR in a mature T cell is a multimeric complex of TCR α and β chains, and CD3 subunits. Functional TCR α and β chains are encoded by genes that result from developmentally controlled somatic rearrangement events. By FACS analysis, we have detected a TCR Vβ8 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 Vβ8 RNA in C1-V13D cells, but no expression of Cβ was detected. Rapid amplification of 3' cDNA ends was used to clone an RNA that was initiated from the leader exon of the Vβ5.1 gene and spliced to the V exon of the Vβ8.2 gene. The putative sequence of the mature 10.8 kDa protein was entirely encoded by the Vβ8.2 exon. RT-PCR analysis confirmed that 97% of the Vβ8 RNA detected in C1-V13D cells was encoded by the Vβ8.2 gene, and only 3% by Vβ8.1 and Vβ8.3 genes. Furthermore, most of the Vβ8.2 RNA was spliced to the leader exon of the Vβ5.1 gene and not to the leader exon of the Vβ8.2gene. The implications of preferential transcription from particular germline TCR genes for repertoire diversity and possible functions for proteins translated from germline TCR Vβ genes are discussed.

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