Characterization of the mouse neurofilament light (NF-L) gene promoter by in vitro transcription.
We have used in vitro transcription to access the basic sequences and factors required for the transcription of the mouse neurofilament light promoter (pNF-L) in the absence of chromatin structure. Deletion from -1.7 to -154 results in little change in NF-L promoter activity using nuclear extracts from either brain (expressing) or liver (non-expressing) tissues. Further deletion to -29 results in a gradual five-fold drop in promoter activity in both extracts. Only replacement of the entire -148 to -29 region results in a drop in NF-L promoter activity to basal levels. Thus, the NF-L promoter differs from the mouse NF heavy (NF-H) and mid-sized (NF-M) promoters in that no specific sequence within the immediate upstream NF-L promoter region (-154 to -29) appears to be responsible for enhancement or brain-specific transcription. We show that the order of strength of the three NF promoters is NF-H>NF-M>NF-L and identify sequences that can increase or reduce transcription when placed in front of heterologous NF promoters. We conclude that the NF-L promoter is a modular, weak and promiscuous promoter whose regulation differs from NF-H or NF-M. Our data suggest that chromatin structure may play an important role in the regulation of the NF-L promoter.
Schwartz
M L
ML
Division of Neuropathology, University of Pennsylvania Medical School, Philadelphia, PA 19104, USA. mlschwar@mail.med.upenn.edu
Hua
Y
Y
Cañete-Soler
R
R
Schlaepfer
W W
WW
D000818:Animals; D002843:Chromatin / Q000235:genetics; D004247:DNA / Q000032:analysis; D004586:Electrophoresis; D017353:Gene Deletion; D005786:Gene Expression Regulation; D066298:In Vitro Techniques; D051379:Mice; D016296:Mutagenesis / Q000502:physiology; D016900:Neurofilament Proteins / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000502:physiology*; D014158:Transcription, Genetic / Q000502:physiology*
D002843:Chromatin; D016900:Neurofilament Proteins; C044882:neurofilament protein L; D004247:DNA