Indirect regulation of presenilins in CREB-mediated transcription.
Presenilins are essential for synaptic function, memory formation, and neuronal survival. Previously, we reported that expression of cAMP response element-binding protein (CREB) target genes is reduced in the cerebral cortex of presenilin (PS) conditional double knock-out (cDKO) mice. To determine whether the reduced expression of the CREB target genes in these mutant mice is due to loss of presenilin directly or secondary to the impaired neuronal activity, we established a sensitive luciferase reporter system to assess direct transcriptional regulation in cultured cells. We first used immortalized PS-deficient mouse embryonic fibroblasts (MEFs), and found that both CREB-mediated transcription and Notch-mediated HES1 transcription are decreased. However, the ubiquitin-C promoter-mediated transcription is also reduced, and among these three reporters, transfection of exogenous PS1 can rescue only the Notch-mediated HES1 transcription. Further Northern analysis revealed transcriptional alterations of Creb, ubiquitin-C, and other housekeeping genes in PS-deficient MEFs, indicating transcriptional dysregulation in these cells. We then used the Cre/loxP system to develop a postnatal PS-deficient cortical neuronal culture. Surprisingly, in these PS-null neurons, CREB-mediated transcription is not significantly decreased, and levels of total and phosphorylated CREB proteins are unchanged as well. Notch-mediated HES1 transcription is markedly reduced, and this reduction can be rescued by exogenous PS1. Together, our findings suggest that CREB-mediated transcription is regulated indirectly by PS in the adult cerebral cortex, and that attenuation of CREB target gene expression in PS cDKO mice is likely due to reduced neuronal activity in these mutant brains.
Watanabe
Hirotaka
H
Center for Neurologic Diseases, Brigham & Women's Hospital Harvard Medical School, Boston, Massachusetts 02115.
Smith
Miriam J
MJ
Center for Neurologic Diseases, Brigham & Women's Hospital Harvard Medical School, Boston, Massachusetts 02115.
Heilig
Elizabeth
E
Center for Human Genetic Research, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts 02115.
Beglopoulos
Vassilios
V
Center for Neurologic Diseases, Brigham & Women's Hospital Harvard Medical School, Boston, Massachusetts 02115.
Kelleher
Raymond J
RJ
Center for Human Genetic Research, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts 02115.
Shen
Jie
J
Center for Neurologic Diseases, Brigham & Women's Hospital Harvard Medical School, Boston, Massachusetts 02115. Electronic address: jshen@rics.bwh.harvard.edu.
D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D017362:Cyclic AMP Response Element-Binding Protein / Q000235:genetics / Q000378:metabolism*; D004622:Embryo, Mammalian / Q000166:cytology / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D051379:Mice; D018345:Mice, Knockout; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D053763:Presenilins / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000502:physiology*; D051880:Receptors, Notch / Q000235:genetics / Q000378:metabolism; D000072056:Transcription Factor HES-1; D014158:Transcription, Genetic / Q000502:physiology*; D025822:Ubiquitin C / Q000235:genetics / Q000378:metabolism
D051792:Basic Helix-Loop-Helix Proteins; D017362:Cyclic AMP Response Element-Binding Protein; D018398:Homeodomain Proteins; D053763:Presenilins; D051880:Receptors, Notch; D000072056:Transcription Factor HES-1; D025822:Ubiquitin C; C489734:Hes1 protein, mouse; C078493:HES1 protein, human