Cloning and functional characterization of the murine caspase-3 gene promoter.
Several studies have shown that the levels of caspase-3 are upregulated under different conditions of apoptosis. Previously, we have shown that activation of T cells through the TCR leads to the upregulation of caspase-3 levels. These findings highlight the importance of regulating the expression of caspase-3 in order to prevent premature cell death. To better understand the regulation of the caspase-3 gene, a portion of the 5'- untranslated region was cloned, sequenced, and characterized. The segment of the 5'-flanking region of the caspase-3 gene was also cloned upstream of a luciferase reporter gene, demonstrating that this fragment contains promoter activity. Higher luciferase expression was found with several of the promoter deletion constructs in Jurkat T cells but not the mouse Neuro-2A neuroblastoma cell line, suggesting the presence of a T-cell-specific regulated region. The importance of these sequences is further supported by the genomic organization of the human and mouse caspase-3 promoter regions. These findings demonstrated that the -2245/+14 region of the caspase-3 promoter shows constitutive levels of expression, and that several regions of the promoter play a role in basal regulation. Finally, some of the conserved transcription factor binding sites identified between the human and mouse promoters appear to play an important role in lymphoid cells.
Sabbagh
Laurent
L
Laboratoire d'Immunologie, Centre de Recherche du CHUM, Campus St.-Luc, Pavillon Edouard-Asselin, 264 Boulevard René Lévesque Est, #1370D, Montreal, Quebec H3C 3J7, Canada.
Bourbonnière
Martin
M
Denis
François
F
Sékaly
Rafick-Pierre
RP
D020121:5' Untranslated Regions; D000818:Animals; D001483:Base Sequence; D053148:Caspase 3; D020169:Caspases / Q000235:genetics*; D045744:Cell Line, Tumor; D003001:Cloning, Molecular; D017930:Genes, Reporter; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic
D020121:5' Untranslated Regions; C505407:CASP3 protein, human; C505409:Casp3 protein, mouse; D053148:Caspase 3; D020169:Caspases