Direct transcriptional induction of Gadd45gamma by Ascl1 during neuronal differentiation.
The basic helix-loop-helix transcription factor Ascl1 plays a critical role in the intrinsic genetic program responsible for neuronal differentiation. Here, we describe a novel model system of P19 embryonic carcinoma cells with doxycycline-inducible expression of Ascl1. Microarray hybridization and real-time PCR showed that these cells demonstrated increased expression of many neuronal proteins in a time- and concentration-dependent manner. Interestingly, the gene encoding the cell cycle regulator Gadd45gamma was increased earliest and to the greatest extent following Ascl1 induction. Here, we provide the first evidence identifying Gadd45gamma as a direct transcriptional target of Ascl1. Transactivation and chromatin immunoprecipitation assays identified two E-box consensus sites within the Gadd45gamma promoter necessary for Ascl1 regulation, and demonstrated that Ascl1 is bound to this region within the Gadd45gamma promoter. Furthermore, we found that overexpression of Gadd45gamma itself is sufficient to initiate some aspects of neuronal differentiation independent of Ascl1.
Huang
Holly S
HS
Neuroscience Graduate Program, University of Michigan, Ann Arbor, MI 48105, USA.
Kubish
Ginger M
GM
Redmond
Tanya M
TM
Turner
David L
DL
Thompson
Robert C
RC
Murphy
Geoffrey G
GG
Uhler
Michael D
MD
D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism*; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology*; D004318:Doxycycline / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D018257:Helix-Loop-Helix Motifs; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D046228:Microarray Analysis; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D018408:Patch-Clamp Techniques; D011401:Promoter Regions, Genetic; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured; D000097642:GADD45 Proteins
D000900:Anti-Bacterial Agents; D051792:Basic Helix-Loop-Helix Proteins; D004318:Doxycycline; D047908:Intracellular Signaling Peptides and Proteins; D000097642:GADD45 Proteins; C499317:Ascl1 protein, mouse
EMBO J. 1990 Nov;9(11):3583-92
2120046
Cell. 1998 Nov 13;95(4):521-30
9827804
Brain Res Dev Brain Res. 1992 May 22;67(1):75-83
1386294
Int J Dev Biol. 1993 Mar;37(1):135-40
8507558
Proc Natl Acad Sci U S A. 2006 Jan 31;103(5):1319-24
16432194
Nature. 1988 Dec 15;336(6200):674-7
3200318
Dev Biol. 1983 Jul;98(1):212-20
6862106
Proc Natl Acad Sci U S A. 1999 Mar 30;96(7):3706-11
10097101
Dev Biol. 2009 Apr 15;328(2):529-40
19389376
Annu Rev Neurosci. 1994;17:267-310
8210176
Biochem Biophys Res Commun. 1996 Jun 25;223(3):691-4
8687458
Nature. 1982 Sep 9;299(5879):165-7
7110336
Science. 1994 Nov 25;266(5189):1376-80
7973727
EMBO J. 1999 Oct 1;18(19):5321-33
10508165
Nature. 1990 Aug 30;346(6287):858-61
2392153
Neurobiology (Bp). 1996;4(4):417-29
9200133
J Neurosci. 1993 Sep;13(9):3986-4000
7690068
Nat Rev Neurosci. 2002 Jul;3(7):517-30
12094208
Int J Oncol. 2001 Apr;18(4):749-57
11251170
J Neurosci. 1983 Nov;3(11):2324-35
6631483
Genes Dev. 1994 Jun 15;8(12):1434-47
7926743
Mol Cell Neurosci. 1999 Oct-Nov;14(4-5):355-69
10588390
Proc Natl Acad Sci U S A. 1996 Oct 15;93(21):11693-8
8876198
Methods Enzymol. 2005;392:186-99
15644182
Annu Rev Neurosci. 2009;32:347-81
19400726
Mech Dev. 2004 Jul;121(7-8):945-58
15210198
Oncogene. 1999 May 6;18(18):2892-900
10362260
BMC Biol. 2008 Mar 31;6:15
18377642
Mol Ther. 2008 Nov;16(11):1873-82
18781144
Development. 2003 Jun;130(11):2505-12
12702663
Nature. 2002 Sep 12;419(6903):135-41
12226657
Proc Natl Acad Sci U S A. 1994 Nov 8;91(23):10800-4
7971965
EMBO J. 2004 Nov 10;23(22):4495-505
15496983
Neuron. 2005 Oct 6;48(1):45-62
16202708
J Cell Sci. 2002 Jan 15;115(Pt 2):329-39
11839785
Proc Natl Acad Sci U S A. 1992 Jun 15;89(12):5547-51
1319065
J Biol Chem. 1999 Mar 26;274(13):8391-404
10085070
Methods Enzymol. 1995;254:20-37
8531687
J Biol Chem. 2005 Apr 22;280(16):16484-98
15722554
Proc Natl Acad Sci U S A. 2000 May 23;97(11):6120-5
10823955
Physiol Res. 2005;54(1):115-22
15717849
J Biol Chem. 2000 Nov 24;275(47):36892-8
10973963
Science. 2009 Feb 20;323(5917):1074-7
19119186
J Comp Neurol. 1988 Jul 15;273(3):344-58
2463277
EMBO J. 2007 Dec 12;26(24):5093-108
18007592
Proc Natl Acad Sci U S A. 2008 May 6;105(18):6566-71
18445651
Science. 1999 Apr 30;284(5415):770-6
10221902
Mech Dev. 2002 Nov;119(1):69-80
12385755
Development. 1992 Jan;114(1):75-87
1576967
Oncogene. 2008 Feb 21;27(9):1263-72
17724474
EMBO J. 1989 Dec 1;8(12):3553-62
2510998
Genes Dev. 1989 Jul;3(7):997-1007
2777079
J Neurosci Res. 1993 Aug 1;35(5):567-76
8377226
Cell. 1993 Nov 5;75(3):463-76
8221886
Dev Cell. 2006 Dec;11(6):831-44
17141158
J Cell Biol. 1989 Apr;108(4):1507-16
2925793
Nature. 2010 Feb 25;463(7284):1035-41
20107439
Cell. 2008 Dec 26;135(7):1201-12
19109892
Development. 2000 Feb;127(4):693-702
10648228
Mol Cell Biol. 2004 Nov;24(21):9517-26
15485919