GREBP, a cGMP-response element-binding protein repressing the transcription of natriuretic peptide receptor 1 (NPR1/GCA).
NPR1/GCA (natriuretic peptide receptor 1/guanylyl cyclase A) expression is controlled by several agents, including ANP (atrial natriuretic peptide). After ANP stimulation, NPR1/GCA down-regulates the transcriptional activity of its gene via a cGMP-dependent mechanism. Because we previously identified a cis-acting element responsible for this cGMP sensitivity, we proceed here to explore novel putative protein binding to cGMP-response element (cGMP-RE). Using the yeast one-hybrid technique with a human kidney cDNA library, we identified a strong positive clone able to bind cGMP-RE. The clone was derived from 1083-bp-long cDNA of a gene of yet unknown function localized on human chromosome 1 (1p33.36). We named this new protein GREBP (for cGMP-response element-binding protein). DNA binding assays showed 18-fold higher cGMP-RE binding capacity than the controls, whereas an electromobility shift assay indicated a specific binding for the cGMP-RE, and chromatin immunoprecipitation confirmed the binding of GREBP to the element under physiological conditions. By acting on cGMP-RE, GREBP inhibited the expression of a luciferase-coupled NPR1 promoter construct. In H295R cells, ANP heightened GREBP expression by 60% after just 3 h of treatment while inhibiting NPR1/GCA expression by 30%. Silencing GREBP with specific small interfering RNA increased the activity of the luciferase-coupled NPR1 promoter and GCA/NPR1 mRNA levels. GREBP is a nuclear protein mainly expressed in the heart. We report here the existence of a human-specific gene that acts as a transcriptional repressor of the NPR1/GCA gene.
Martel
Guy
G
Research Centre, Centre Hospitalier de l'Université de Montréal (CRCHUM), Montréal, Québec H1W 4A4, Canada.
Hamet
Pavel
P
Tremblay
Johanne
J
D001483:Base Sequence; D002352:Carrier Proteins / Q000378:metabolism*; D002878:Chromosomes, Human, Pair 1; D003001:Cloning, Molecular; D006152:Cyclic GMP / Q000378:metabolism; D017931:DNA Primers / Q000737:chemistry / Q000302:isolation & purification; D016373:DNA, Antisense / Q000235:genetics; D024202:Electrophoretic Mobility Shift Assay; D005786:Gene Expression Regulation; D006367:HeLa Cells; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism*; D007668:Kidney; D008969:Molecular Sequence Data; D010957:Plasmids; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics; D017461:Receptors, Atrial Natriuretic Factor / Q000235:genetics*; D014158:Transcription, Genetic
D002352:Carrier Proteins; D017931:DNA Primers; D016373:DNA, Antisense; D047908:Intracellular Signaling Peptides and Proteins; D012333:RNA, Messenger; C022994:cyclic GMP-binding protein; D017461:Receptors, Atrial Natriuretic Factor; C091715:atrial natriuretic factor receptor A; D006152:Cyclic GMP
Clin Genet. 2007 Mar;71(3):238-44
17309646
Methods. 1999 Dec;19(4):506-20
10581150
J Mol Biol. 1999 Dec 17;294(5):1351-62
10600390
Am J Physiol Renal Physiol. 2005 Dec;289(6):F1273-80
16033922
Methods. 2002 Jan;26(1):37-47
12054903
J Biol Chem. 1991 Dec 5;266(34):23060-7
1660465
J Cyclic Nucleotide Res. 1978 Aug;4(4):281-90
82561
Hypertension. 2003 Mar;41(3 Pt 2):730-6
12623988
Biochem Biophys Res Commun. 1990 Apr 30;168(2):863-70
2139780
Nucleic Acids Res. 1987 Oct 26;15(20):8125-48
3313277
Science. 1983 Jul 1;221(4605):71-3
6857267
J Biol Chem. 1998 May 29;273(22):13444-51
9593677
Proc Natl Acad Sci U S A. 2009 May 5;106(18):7273-80
19351897
Mol Pharmacol. 2004 Feb;65(2):319-25
14742673
Hypertension. 2004 Jun;43(6):1270-8
15096467
Nucleic Acids Res. 1999 Sep 15;27(18):3728-35
10471743
J Immunol. 2000 Jul 1;165(1):175-81
10861050
Annu Rev Physiol. 1990;52:747-59
2139559
Biochem J. 2002 Aug 1;365(Pt 3):561-75
12006103
J Biol Chem. 2003 Dec 5;278(49):48617-26
12970365
Hypertension. 1992 Jun;19(6 Pt 2):758-61
1534317
Biochem Biophys Res Commun. 2007 Nov 23;363(3):674-9
17904522
J Clin Invest. 1990 Nov;86(5):1690-7
2173726
Cell. 1992 Oct 2;71(1):1-4
1356629
Endocrinology. 1991 Nov;129(5):2311-7
1657570
J Biol Chem. 1992 Apr 15;267(11):7624-9
1348507
Peptides. 2005 Jun;26(6):1009-23
15911069
Proc Natl Acad Sci U S A. 1984 Dec;81(23):7661-4
6150486
Endocrinology. 1984 Nov;115(5):2026-8
6092045
Biochem Biophys Res Commun. 1989 Sep 29;163(3):1435-43
2551288
Mol Cell Biochem. 2010 Jan;334(1-2):53-65
19937369
J Biol Chem. 1995 Oct 20;270(42):24891-7
7559613
J Cell Sci. 2000 Jul;113 ( Pt 14):2527-33
10862710
Biochem Biophys Res Commun. 1984 Sep 17;123(2):515-27
6091631
Am J Physiol Renal Physiol. 2001 Mar;280(3):F466-73
11181408
J Biol Chem. 1998 Feb 13;273(7):4220-6
9461619
EMBO J. 1999 Aug 16;18(16):4549-59
10449420
FEBS Lett. 2007 Sep 4;581(22):4325-32
17716668
Nature. 1988 Mar 3;332(6159):78-81
2964562
Trends Biochem Sci. 1995 Nov;20(11):465-70
8578590
Biochemistry. 1999 Dec 21;38(51):16941-51
10606529
J Biol Chem. 2005 Oct 21;280(42):35184-94
16105833
J Biol Chem. 1989 Feb 25;264(6):3043-6
2536732
Life Sci. 1981 Jan 5;28(1):89-94
7219045
J Biol Chem. 2002 Aug 30;277(35):32003-14
12082086
Protein Eng Des Sel. 2004 Nov;17(11):809-19
15576381
Trends Genet. 1996 Sep;12(9):340-1
8855661
Hypertension. 2003 Mar;41(3 Pt 2):675-81
12623978
Eur J Biochem. 2003 May;270(10):2174-85
12752437
Am J Physiol Renal Physiol. 2000 Jul;279(1):F122-9
10894794
FEBS Lett. 1986 Jan 6;194(2):210-4
2866981
Nat Cell Biol. 2008 Feb;10(2):202-10
18193036
Nat Rev Mol Cell Biol. 2002 Jan;3(1):30-40
11823796
EMBO J. 1997 Apr 15;16(8):1865-75
9155013
Mol Pharmacol. 2002 Oct;62(4):927-35
12237340
J Mol Cell Cardiol. 1997 Sep;29(9):2375-82
9299361
Cell. 1986 Aug 29;46(5):659-67
3488815
J Cell Biochem. 2005 May 15;95(2):217-25
15770648
J Biol Chem. 2003 Mar 14;278(11):9472-80
12524425
Nat Rev Mol Cell Biol. 2002 Sep;3(9):710-8
12209131
Hypertension. 1995 Apr;25(4 Pt 2):711-4
7536712
Nature. 2006 Mar 9;440(7081):242-5
16525476
J Biol Chem. 1994 Dec 9;269(49):30741-4
7982997