NeuroD Factors Discriminate Mineralocorticoid From Glucocorticoid Receptor DNA Binding in the Male Rat Brain.
In the limbic brain, mineralocorticoid receptors (MRs) and glucocorticoid receptors (GRs) both function as receptors for the naturally occurring glucocorticoids (corticosterone/cortisol) but mediate distinct effects on cellular physiology via transcriptional mechanisms. The transcriptional basis for specificity of these MR- vs GR-mediated effects is unknown. To address this conundrum, we have identified the extent of MR/GR DNA-binding selectivity in the rat hippocampus using chromatin immunoprecipitation followed by sequencing. We found 918 and 1450 nonoverlapping binding sites for MR and GR, respectively. Furthermore, 475 loci were co-occupied by MR and GR. De novo motif analysis resulted in a similar binding motif for both receptors at 100% of the target loci, which matched the known glucocorticoid response element (GRE). In addition, the Atoh/NeuroD consensus sequence was found in co-occurrence with all MR-specific binding sites but was absent for GR-specific or MR-GR overlapping sites. Basic helix-loop-helix family members Neurod1, Neurod2, and Neurod6 showed hippocampal expression and were hypothesized to bind the Atoh motif. Neurod2 was detected at rat hippocampal MR binding sites but not at GR-exclusive sites. All three NeuroD transcription factors acted as DNA-binding-dependent coactivators for both MR and GR in reporter assays in heterologous HEK293 cells, likely via indirect interactions with the receptors. In conclusion, a NeuroD family member binding to an additional motif near the GRE seems to drive specificity for MR over GR binding at hippocampal binding sites.
Department of Medicine, Division of Endocrinology, Leiden University Medical Center, 2300 RC, Leiden, The Netherlands.|Department of Cognitive Neuroscience, Radboud University Medical Center, 6500 HB, Nijmegen, The Netherlands.|Donders Institute for Brain, Cognition, and Behaviour, Radboud University, 6525 EN, Nijmegen, The Netherlands.
Lisa T C M
LTCM
van Weert
Department of Medicine, Division of Endocrinology, Leiden University Medical Center, 2300 RC, Leiden, The Netherlands.
Jacobus C
JC
Buurstede
Department of Radiology, Division of Image Processing, Leiden University Medical Center, 2300 RC, Leiden, The Netherlands.|Delft Bioinformatics Laboratory, Delft University of Technology, 2628 CD, Delft, The Netherlands.
Ahmed
A
Mahfouz
Department of Medicine, Division of Endocrinology, Leiden University Medical Center, 2300 RC, Leiden, The Netherlands.
Pamela S M
PSM
Braakhuis
Division of Medical Pharmacology, Leiden/Amsterdam Center for Drug Research, 2300 RC, Leiden, The Netherlands.
J Annelies E
JAE
Polman
Department of Medicine, Division of Endocrinology, Leiden University Medical Center, 2300 RC, Leiden, The Netherlands.
Hetty C M
HCM
Sips
Department of Cognitive Neuroscience, Radboud University Medical Center, 6500 HB, Nijmegen, The Netherlands.|Donders Institute for Brain, Cognition, and Behaviour, Radboud University, 6525 EN, Nijmegen, The Netherlands.
Benno
B
Roozendaal
Department of Human Genetics, Leiden University Medical Center, 2300 RC, Leiden, The Netherlands.
Judit
J
Balog
Department of Medicine, Division of Endocrinology, Leiden University Medical Center, 2300 RC, Leiden, The Netherlands.|Division of Medical Pharmacology, Leiden/Amsterdam Center for Drug Research, 2300 RC, Leiden, The Netherlands.
E Ronald
ER
de Kloet
Division of Medical Pharmacology, Leiden/Amsterdam Center for Drug Research, 2300 RC, Leiden, The Netherlands.
Nicole A
NA
Datson
Department of Medicine, Division of Endocrinology, Leiden University Medical Center, 2300 RC, Leiden, The Netherlands.
Onno C
OC
Meijer
D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000378:metabolism / Q000502:physiology*; D001665:Binding Sites; D001921:Brain / Q000378:metabolism*; D004247:DNA / Q000378:metabolism; D005260:Female; D057809:HEK293 Cells; D006801:Humans; D008297:Male; D009479:Neuropeptides / Q000378:metabolism / Q000502:physiology*; D011485:Protein Binding; D051381:Rats; D017207:Rats, Sprague-Dawley; D011965:Receptors, Glucocorticoid / Q000378:metabolism*; D018161:Receptors, Mineralocorticoid / Q000378:metabolism*; D020218:Response Elements / Q000235:genetics*
D051792:Basic Helix-Loop-Helix Proteins; D004247:DNA; D009479:Neuropeptides; D011965:Receptors, Glucocorticoid; D018161:Receptors, Mineralocorticoid; C490924:Neurod2 protein, rat