Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
4330526 | Brain Research | 2007 | 12 Pages |
Abstract
The expression of glutamate carboxypeptidase II (GCP II) is reduced in selective brain regions in schizophrenic patients. To investigate transcriptional mechanisms regulating the human GCP II gene, a 3460Â bp DNA fragment comprised of the proximal 3228Â bp of 5â² untranscribed sequence and first 232Â bp of 5â² UTR portion of this gene was cloned into the mammalian luciferase reporter gene vector pGL3-Basic. Transfection assays in human astrocyte-derived SVG and human prostate tumor-derived LNCaP cells demonstrated that constructs with 3460, 1590 and 761Â bp portions of 5â² region of human GCP II gene were able to drive the luciferase reporter gene. Additional deletion constructs showed that in the SVG cell line, constructs with 511 and 411Â bp of GCP II gene fragments yielded highest transcriptional activity, with declining activity upon further removal of 5â² sequences. 15Â bp of the promoter 5â² to a 225Â bp GCP II fragment were essential for luciferase expression. Thus, in the SVG cells, the proximal 240Â bp of the human GCP II promoter (232Â bp of the 5â² UTR and 8Â bp of 5â² untranscribed sequences) may represent the core promoter. Further, while a LyF-1 site lies within and overlaps a transcription start site in the 15Â bp sequence, site-directed mutagenesis shows that LyF-1 is not the transcription initiator for the “TATA and CAAT” box lacking GCP II gene in the SVG cells. Finally, pattern differences in GCP II gene promoter expression in SVG and LNCaP cells suggest that sequences beyond 240Â bp may be important for tissue-specific GCP II expression.
Related Topics
Life Sciences
Neuroscience
Neuroscience (General)
Authors
Liqun Han, Dona Lee Wong, Guochuan Tsai, Zhichun Jiang, Joseph T. Coyle,