Cloning of the murine CD5 promoter and its tissue-specific regulation.
A genomic clone containing 1700 bp of the 5'-flanking region and first exon of the murine CD5 gene was isolated by screening a NIH 3T3 fibroblast genomic library with the previously characterized murine CD5 cDNA. The CD5 5'-flanking region lacks a consensus TATA box but contains Sp1, AP-1, IgH muE2, SV40 enhancer core, CCAAT, TCF2 alpha/PEA3/ets, and C/EBP motifs. Transcription is initiated from multiple sites upstream of an Inr-like sequence. When linked to the CAT reporter gene, the CD5 5'-flanking region was an active promoter in transient transfection assays using the EL4 (T), PD36 (pre-B) and M12 (B) cell lines, but was inactive in NIH 3T3 fibroblasts. This pattern of lymphoid-specific expression reflects the pattern of in vivo CD5 expression. Successive 5' to 3' deletions of the CD5 promoter/CAT reporter construct were transfected into T and B cells revealing a succession of positive and negative regulatory elements until promoter activity was eliminated at position -27. This result identified sequences from -125 to -27 (which contain the Inr, IgH muE2, SV40 enhancer core, and AP-1 sites) as sufficient for tissue-specific promotion. Isolation and characterization of the CD5 promoter represents an initial step in elucidating the control over tissue-specific expression of the murine CD5 gene.
Weichert
T R
TR
Department of Microbiology, Michigan State University, East Lansing 48824, USA.
Schwartz
R C
RC
D000818:Animals; D015703:Antigens, CD / Q000235:genetics*; D001483:Base Sequence; D018956:CD5 Antigens; D002460:Cell Line; D003001:Cloning, Molecular / Q000379:methods*; D005091:Exons; D005786:Gene Expression Regulation / Q000276:immunology*; D051379:Mice; D008969:Molecular Sequence Data; D009928:Organ Specificity / Q000235:genetics / Q000276:immunology; D010957:Plasmids; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012045:Regulatory Sequences, Nucleic Acid
D015703:Antigens, CD; D018956:CD5 Antigens