Human CD3-CD16+ natural killer cells express the hGATA-3 T cell transcription factor and an unrearranged 2.3-kb TcR delta transcript.
In this study we analyzed the T cell receptor(TcR) delta transcripts expressed by CD3-CD16+ cells and we investigated whether these cells expressed the hGATA-3 T cell transcription factor and the recombination-activating gene (RAG)-1. Multiple TcR delta transcripts deriving from an unrearranged TcR delta gene were detected in both polyclonal and clonal CD3-CD16+ natural killer(NK) cell lines. Two unrearranged TcR delta transcripts had a size similar to that of the functional TcR delta mRNA (2.3 and 1.3 kb) found in TcR gamma/delta+ T lymphocytes. Sequence analysis of nine different 2.3-kb cDNA clones obtained from NK-derived polyA+ RNA confirmed that they corresponded to an unrearranged TcR delta gene. These cDNA were 2343 bp long and their transcription initiation site was located 814 bp upstream from the J delta 1 segment. The sequence located upstream of the J delta 1 segment corresponded to the previously reported germ-line sequence. The J delta 1 segment was correctly spliced to C delta; in addition the four C delta exons were found to be already assembled. Two polyadenylation sites were present in the fourth C delta exon. However, only that located at the 3' end appeared to be utilized in the 2.3-kb cDNA. The expression of hGATA-3, a T cell-specific factor known to be involved in the regulation of the transcription of TcR delta locus, was analyzed by Northern blot, in cultured NK cell population and clones (but not in freshly derived cell populations). All NK clones and cell lines studied were found to express hGATA-3-specific mRNA, suggesting that hGATA-3 may be involved in the regulation of the unrearranged TcR delta gene expression in NK cells. Finally, no transcription of the RAG-1 gene could be detected in all NK cell lines or clones analyzed.
Lab. Farmacologia, Istituto Nazionale per la Ricerca sul Cancro, Genova, Italy.
R
R
Biassoni
S
S
Verdiani
A
A
Cambiaggi
P H
PH
Romeo
S
S
Ferrini
L
L
Moretta
D001483:Base Sequence; D017252:CD3 Complex / Q000032:analysis*; D002460:Cell Line; D004247:DNA / Q000302:isolation & purification; D004268:DNA-Binding Proteins / Q000032:analysis*; D050990:GATA3 Transcription Factor; D016694:Gene Rearrangement, delta-Chain T-Cell Antigen Receptor; D018398:Homeodomain Proteins; D006801:Humans; D007694:Killer Cells, Natural / Q000276:immunology*; D008969:Molecular Sequence Data; D011506:Proteins / Q000235:genetics; D016692:Receptors, Antigen, T-Cell, gamma-delta / Q000235:genetics*; D017452:Receptors, IgG / Q000032:analysis*; D015534:Trans-Activators / Q000032:analysis*; D014157:Transcription Factors / Q000032:analysis*; D014158:Transcription, Genetic
D017252:CD3 Complex; D004268:DNA-Binding Proteins; D050990:GATA3 Transcription Factor; C494749:GATA3 protein, human; D018398:Homeodomain Proteins; D011506:Proteins; D016692:Receptors, Antigen, T-Cell, gamma-delta; D017452:Receptors, IgG; D015534:Trans-Activators; D014157:Transcription Factors; C064658:RAG-1 protein; D004247:DNA