Transcriptional Regulation of Gfap

Transcriptional regulation of the glial fibrillary acidic protein gene (GFAP) is of interest because of its astrocyte specificity and its up-regulation in response to CNS injuries. We have used a transgenic approach instead of cell transfection to identify promoter elements of the human GFAP gene, since previous observations show that GFAP transcription is regulated differently in transfected cultured cells than in the mouse. We previously showed that block mutation of enhancer regions spanning bp 1488 to -1434 (the C1.1 segment) and -1443 to -1399 (C1.2) resulted in altered patterns of expression and loss of astrocyte specificity, respectively. This analysis has now been extended upstream to bp -1612 to -1489 (the B region), which previously has been shown especially important for expression. Block mutation of each of 4 contiguous sequences which together span the B region each decreased the level of transgene activity by at least 50%, indicating that multiple sites contribute to the transcriptional activity in a cooperative manner. Several of the block mutations also altered the brain region pattern of expression, astrocyte specificity and/or the developmental time course. Transgenes were then analyzed in which mutations were limited to specific transcription factor binding sites in each of the 4 B block segments as well as in C1.1 and C1.2. Whereas mutation of the conserved consensus AP-1 site unexpectedly had no discernible effect on transgene expression; NFI, SP1, STAT3 and NF-κB were identified as having important roles in regulating the strength of GFAP promoter activity and/or its astrocyte specificity. INTRODUCTION Transcriptional regulation of the glial fibrillary acidic protein (GFAP) gene has attracted interest due to its astrocyte specificity and its upregulation in response to