Multiple cis-acting elements are required for proper transcription of the mouse V delta 1 T cell receptor promoter.
To gain insight into the developmentally regulated expression of the mouse TCR V delta-gene segments, we have investigated the role of the 5' promoter region of the V delta 1-gene. Transient transfection assays showed that a construct encompassing 267 nucleotides upstream from the mapped transcriptional start site was capable of driving promoter activity when transfected into V delta 1+ T cells. The inclusion of an additional 459-bp 5' segment to this construct did not affect promoter activity. However, a deletion of 222 5' nucleotides from the same construct dramatically decreased promoter activity. In vivo genomic footprinting localized several protein-DNA interactions to the stretch of DNA shown to have transcriptional activity. A computer analysis revealed that the segments of DNA participating in these protein-DNA interactions were identical to the previously described cyclic AMP response element (CRE), E box, and leukemia virus E26 cis-acting elements. Transient transfection assays performed with -267 bp constructs containing mutations at each of the localized cis-acting elements revealed that the CRE, E box, and Ets elements work together in driving promoter activity and that the CRE and Ets elements are the most important for driving transcription. Gel mobility shift analyses showed that each of these cis-acting elements is capable of binding specific nuclear factors present in V delta 1-expressing cells. These data indicate that multiple transcription factors acting in concert are responsible for V delta 1 gene expression.
Punturieri
A
A
Biological Resources Branch, National Institute of Allergy and Infectious Diseases, Bethesda, MD 20892.
Shirakata
Y
Y
Bovolenta
C
C
Kikuchi
G
G
Coligan
J E
JE
D000818:Animals; D001483:Base Sequence; D002874:Chromosome Mapping; D004252:DNA Mutational Analysis; D004591:Electrophoresis, Polyacrylamide Gel; D005809:Genes, Regulator; D007135:Immunoglobulin Variable Region / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D009154:Mutation; D011401:Promoter Regions, Genetic; D016692:Receptors, Antigen, T-Cell, gamma-delta / Q000235:genetics*; D014158:Transcription, Genetic
D007135:Immunoglobulin Variable Region; D016692:Receptors, Antigen, T-Cell, gamma-delta