Differential roles for ETS, CREB, and EGR binding sites in mediating VEGF receptor 1 expression in vivo.
Vascular endothelial growth factor receptor 1 (VEGFR1) is a marker for endothelial-specific gene expression. We previously reported that the human VEGFR1 promoter (between -748 and +284) contains information for expression in the intact endothelium of transgenic mice. The objective of this study was to dissect the cis-regulatory elements underlying VEGFR1 promoter activity in vitro and in vivo. In primary endothelial cells, binding sites for E74-like factor 1 (ELF-1; between -49 and -52), cyclic adenosine monophosphate response element binding (CREB; between -74 and -81), and early growth response factor 1/3 (EGR-1/3; between -16 to -25) were shown to play a positive role in gene transcription, whereas a putative E26 transformation-specificsequence (ETS) motif between -36 and -39 had a net negative effect on promoter activity. When targeted to the Hprt locus of mice, mutations of the ELF-1 binding site and the CRE element reduced promoter activity in the embryonic vasculature and resulted in a virtual loss of expression in adult endothelium. Postnatally, the EGR binding site mutant displayed significantly reduced promoter activity in a subset of vascular beds. In contrast, mutation of the -39 ETS site resulted in increased LacZ staining in multiple vascular beds. Together, these results provide new insights into the transcriptional regulatory mechanisms of VEGFR1.
Jin
Enjing
E
The Center for Vascular Biology Research and Division of Molecular and Vascular Medicine, Beth Israel Deaconess Medical Center, Boston, MA, USA.
Liu
Ju
J
Suehiro
Jun-Ichi
J
Yuan
Lei
L
Okada
Yoshiaki
Y
Nikolova-Krstevski
Vesna
V
Yano
Kiichiro
K
Janes
Lauren
L
Beeler
David
D
Spokes
Katherine C
KC
Li
Dan
D
Regan
Erzsébet
E
Shih
Shou-Ching
SC
Oettgen
Peter
P
Minami
Takashi
T
Aird
William C
WC
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics / Q000502:physiology; D050882:CREB-Binding Protein / Q000378:metabolism*; D002478:Cells, Cultured; D051766:Early Growth Response Protein 1 / Q000378:metabolism; D051776:Early Growth Response Protein 3 / Q000378:metabolism; D051765:Early Growth Response Transcription Factors / Q000378:metabolism*; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism; D005260:Female; D005786:Gene Expression Regulation / Q000502:physiology; D006801:Humans; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D009154:Mutation; D009687:Nuclear Proteins / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011485:Protein Binding; D012689:Sequence Homology, Nucleic Acid; D014157:Transcription Factors / Q000378:metabolism*; D040281:Vascular Endothelial Growth Factor Receptor-1 / Q000235:genetics*
C497325:EGR1 protein, human; C079945:EGR3 protein, human; C512897:ELF1 protein, human; D051766:Early Growth Response Protein 1; D051765:Early Growth Response Transcription Factors; D009687:Nuclear Proteins; D014157:Transcription Factors; D051776:Early Growth Response Protein 3; D050882:CREB-Binding Protein; D040281:Vascular Endothelial Growth Factor Receptor-1