Cloning and characterization of the human and rabbit NUDEL-oligopeptidase promoters and their negative regulation.
NUDEL-oligopeptidase is a cytosolic cysteine peptidase, active towards oligopeptides and involved in the conversion and inactivation of a number of bioactive peptides. This protein interacts with neuronal proteins and is essential for brain development and cortical organization during embryogenesis. In this study, 5'-flanking sequences of the human and rabbit NUDEL-oligopeptidase gene were cloned into the pGL3 reporter gene vector and the promoter activity of the full-length fragment and deletions series was measured in transient transfection assays using two different cell lines, namely, C6 rat glioma and NH15 human neuroblastoma. Overall, a very similar pattern of promoter activity was obtained for both rabbit and human NUDEL-oligopeptidase promoter sequences, and their respective serial deletion constructs upon transient transfection into these cell lines. The only exception was for the longest rabbit upstream sequence that displayed about 1.8-fold higher luciferase expression upon transfection into NH15 neuronal cells than that observed upon transfection into C6 glioma cells. On the other hand, no significant difference was observed for the human longest sequence. These results are in good agreement with the expression pattern of NUDEL-oligopeptidase in human and rabbit tissues.
Guerreiro
Juliano R
JR
Laboratory for Applied Toxinology, Butantan Institute, São Paulo, 05503-900, Brazil.
Winnischofer
Sheila M B
SM
Bastos
Marta F
MF
Portaro
Fernanda C V
FC
Sogayar
Mari C
MC
de Camargo
Antonio C M
AC
Hayashi
Mirian A F
MA
D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D002352:Carrier Proteins / Q000235:genetics*; D045744:Cell Line, Tumor; D002490:Central Nervous System / Q000378:metabolism; D003001:Cloning, Molecular; D005786:Gene Expression Regulation; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D008156:Luciferases; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011817:Rabbits; D051381:Rats; D017422:Sequence Analysis, DNA
D002352:Carrier Proteins; C421108:NDEL1 protein, human; D008156:Luciferases