VE-statin/egfl7 expression in endothelial cells is regulated by a distal enhancer and a proximal promoter under the direct control of Erg and GATA-2.
Angiogenesis is the process by which new blood vessels arise from existing ones by the budding out of endothelial cell capillaries from the luminal side of blood vessels. Blood vessel formation is essential for organ development during embryogenesis and is associated with several physiological and pathological processes, such as wound healing and tumor development. The VE-statin/egfl7 gene is specifically expressed in endothelial cells during embryonic development and in the adult. We studied here the regulatory mechanisms that control this tissue-specific expression. RT-qPCR analyses showed that the specificity of expression of VE-statin/egfl7 in endothelial cells is not shared with its closest neighbor genes notch1 and agpat2 on the mouse chromosome 2. Chromatin-immunoprecipitation analysis of histone modifications at the VE-statin/egfl7 locus showed that the chromatin is specifically opened in endothelial cells, but not in fibroblasts at the transcription start sites. A 13 kb genomic fragment of promoter was cloned and analyzed by gene reporter assays which showed that two conserved regions are important for the specific expression of VE-statin/egfl7 in endothelial cells; a -8409/-7563 enhancer and the -252/+38 region encompassing the exon-1b transcription start site. The latter contains essential GATA and ETS-binding sites, as assessed by linker-scanning analysis and site-directed mutagenesis. An analysis of expression of the ETS and GATA transcription factors showed that Erg, Fli-1 and GATA-2 are the most highly expressed factors in endothelial cells. Erg and GATA-2 directly control the expression of the endogenous VE-statin/egfl7 while Fli-1 probably exerts an indirect control, as assessed by RNA interference and chromatin immunoprecipitation. This first detailed analysis of the mechanisms that govern the expression of the VE-statin/egfl7 gene in endothelial cells pinpoints the specific importance of ETS and GATA factors in the specific regulation of genes in this cell lineage.
Le Bras
Alexandra
A
CNRS, Institut de Biologie de Lille, UMR 8161, Equipe labellisée La Ligue, Lille, France.
Samson
Chantal
C
Trentini
Matteo
M
Caetano
Bertrand
B
Lelievre
Etienne
E
Mattot
Virginie
V
Beermann
Friedrich
F
Soncin
Fabrice
F
D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D002135:Calcium-Binding Proteins; D033481:Chromosomes, Mammalian / Q000235:genetics; D066255:EGF Family of Proteins; D042783:Endothelial Cells / Q000378:metabolism*; D004742:Enhancer Elements, Genetic / Q000235:genetics*; D050989:GATA2 Transcription Factor / Q000378:metabolism*; D017353:Gene Deletion; D005786:Gene Expression Regulation / Q000235:genetics*; D056426:Genetic Loci / Q000235:genetics; D006801:Humans; D051379:Mice; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011506:Proteins / Q000235:genetics* / Q000378:metabolism; D051789:Proto-Oncogene Protein c-fli-1 / Q000378:metabolism*; D014158:Transcription, Genetic
D002135:Calcium-Binding Proteins; D066255:EGF Family of Proteins; C479627:Egfl7 protein, mouse; C497426:Fli1 protein, mouse; D050989:GATA2 Transcription Factor; C494712:Gata2 protein, mouse; D011506:Proteins; D051789:Proto-Oncogene Protein c-fli-1
Nature. 1995 Oct 19;377(6550):639-42
7566177
J Biol Chem. 2003 Mar 7;278(10):8385-94
12511565
J Biol Chem. 2007 May 11;282(19):14665-74
17347142
J Biol Chem. 1992 Jan 15;267(2):1279-85
1370462
Oncogene. 2000 May 11;19(20):2438-46
10828886
Nat Immunol. 2008 Jul;9(7):810-9
18500345
Cell. 2008 Dec 12;135(6):1053-64
19070576
Blood. 2008 Apr 1;111(7):3498-506
18195090
Nature. 2004 Apr 15;428(6984):754-8
15085134
EMBO J. 1997 Jul 16;16(14):4374-83
9250681
Dev Cell. 2009 Feb;16(2):180-95
19217421
Blood. 2008 Sep 15;112(6):2336-9
18519813
Development. 2007 Aug;134(16):2913-23
17626061
J Biol Chem. 1998 May 8;273(19):11737-44
9565596
Development. 1992 Jul;115(3):737-44
1425352
Development. 1989 Oct;107(2):265-74
2483681
Proc Natl Acad Sci U S A. 1985 Apr;82(8):2230-4
3857577
J Biol Chem. 2004 Aug 13;279(33):35087-100
15180995
Circ Res. 2007 Jun 22;100(12):1712-22
17495228
J Biol Chem. 1996 Nov 29;271(48):30823-8
8940064
Nat Cell Biol. 2009 Jul;11(7):873-80
19503073
Proc Natl Acad Sci U S A. 2009 Jan 20;106(3):814-9
19129488
Proc Natl Acad Sci U S A. 1994 Jul 19;91(15):7291-5
8041783
J Biol Chem. 2002 Jul 12;277(28):25143-51
11991951
Nature. 1995 Oct 19;377(6550):635-8
7566176
Blood. 2000 Nov 1;96(9):3078-85
11049987
Science. 1993 Jul 9;261(5118):203-6
8327891
Mol Cell. 2005 Sep 2;19(5):631-42
16137620
J Biol Chem. 1997 Aug 8;272(32):20299-305
9242711
Circ Res. 1999 May 28;84(10):1177-85
10347092
Gene. 1989 Apr 15;77(1):51-9
2744487
Blood. 2006 Jun 15;107(12):4737-45
16484587
Mol Cell Biol. 1994 Feb;14(2):999-1008
7507210
Gene Expr Patterns. 2008 Jul;8(6):389-96
18556249
RNA. 2006 Jul;12(7):1161-7
16738409
Blood. 2009 Jul 30;114(5):1123-30
19411629
EMBO J. 2001 Sep 17;20(18):5139-52
11566878
Mol Cell Biol. 2007 Apr;27(7):2687-97
17242194
EMBO J. 2003 Nov 3;22(21):5700-11
14592969
Dev Cell. 2008 Aug;15(2):261-71
18694565
Blood. 1999 Jun 15;93(12):4284-92
10361126
Immunity. 1998 Oct;9(4):555-63
9806641
Mech Dev. 1995 Mar;50(1):17-28
7605748
J Biol Chem. 1995 Nov 17;270(46):27948-53
7499271
Curr Opin Genet Dev. 2009 Dec;19(6):565-70
19913403
Blood. 1995 Sep 1;86(5):1828-35
7655012
Biochim Biophys Acta. 2007 Jun;1775(2):298-312
17572301
FASEB J. 1999 Feb;13(2):377-86
9973326
J Biol Chem. 1995 Jun 23;270(25):15320-6
7541039
Proc Natl Acad Sci U S A. 1997 Apr 1;94(7):3058-63
9096345
Arterioscler Thromb Vasc Biol. 1997 Aug;17(8):1599-604
9301641
Int J Dev Biol. 1998 May;42(4):561-72
9694627
Hum Mol Genet. 2005 Mar 1;14(5):595-601
15649946
Int J Biochem Cell Biol. 2001 Apr;33(4):391-407
11312108
Mol Cell Biol. 2005 Feb;25(4):1458-74
15684396
Development. 2008 Dec;135(24):3989-93
18987025
J Biol Chem. 2005 Jul 1;280(26):24824-38
15870070
Blood. 2006 Aug 15;108(4):1251-9
16627757
Circ Res. 2008 Nov 7;103(10):1147-54
18832752
Genes Dev. 2000 Jun 1;14(11):1343-52
10837027
EMBO J. 2008 Jun 18;27(12):1658-70
18497746
Mol Cell Biol. 1990 Sep;10(9):4854-62
2388628
Cell Metab. 2009 Feb;9(2):165-76
19187773
Proc Natl Acad Sci U S A. 2007 Nov 6;104(45):17692-7
17962413
J Biol Chem. 1998 Mar 20;273(12):6750-5
9506975
Nucleic Acids Res. 2004 Oct 21;32(18):5693-702
15498926