Transcriptional regulation of the rat PLP promoter in primary cultures of oligodendrocytes.
Proteolipid protein (PLP) is the major intrinsic membrane protein of CNS myelin and is expressed in oligodendrocytes as part of a coordinate program of myelin-specific gene activation. In order to identify the DNA sequences and proteins involved in the regulation of PLP transcription, we have analyzed the 5' flanking sequences of the rat PLP gene by transient transfections into primary cultures of developing oligodendrocytes, the glial tumor line, C6, and L cells. High levels of expression of the CAT reporter gene in oligodendrocytes and C6 cells were obtained with constructs containing both 4270 and 225 bp of PLP promoter. A fusion construct containing 1061 bp of the PLP promoter, however, showed two-fold lower CAT expression. In addition, the activity of these promoter fusion constructs in oligodendrocytes was 2.5-4.6 higher than that observed in C6 cells, while very little expression was found in L cells. These data suggest that 225 bp of PLP promoter is sufficient for oligodendrocyte-specific regulation of PLP expression. Furthermore, both positive and negative elements within the PLP promoter are involved in this process. Finally, primary cultures of developing oligodendrocytes are a useful model system for the analysis of myelin-specific gene activation.
Cambi
F
F
Department of Neurology, Thomas Jefferson University, Philadelphia, PA 19107.
Kamholz
J
J
D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D009185:Myelin Proteins / Q000235:genetics*; D018991:Myelin Proteolipid Protein; D009836:Oligodendroglia / Q000502:physiology*; D011401:Promoter Regions, Genetic; D051381:Rats; D014158:Transcription, Genetic
D009185:Myelin Proteins; D018991:Myelin Proteolipid Protein; D015500:Chloramphenicol O-Acetyltransferase