In vivo analysis of key elements within the renin regulatory region.
Renin is responsible for initiating the enzymatic cascade that results in the production of angiotensin II, the major effector molecule of the renin-angiotensin system (RAS). Extensive information on the regulatory region of the renin gene has been derived by transient transfection studies in vitro, particularly using the As4.1 cell line. To verify key factors within the regulatory region of renin in vivo, homologous recombination was used to introduce a green fluorescent protein (GFP) cassette into exon one of the renin gene contained within a 240 kb bacterial artificial chromosome (BAC) to create a construct that has GFP expression controlled by the renin regulatory region (RenGFP BAC). Within the regulatory region of the RenGFP BAC construct we independently deleted the enhancer, as well as mutated the HOX-PBX site within the proximal promoter element. Transgenic lines were generated for each of these BAC constructs and GFP expression was analyzed throughout a spectrum of tissues positive for renin expression including the kidney, adrenal gland, gonadal artery, and submandibular gland. The results described within this manuscript support the interpretation that the renin enhancer is critical for regulating baseline expression where as the Hox/Pbx site is important for the tissue specificity of renin expression.
Glenn
Sean T
ST
Department of Molecular and Cellular Biology, Roswell Park Cancer Institute, Buffalo, New York 14263-0001, USA.
Jones
Craig A
CA
Pan
Li
L
Gross
Kenneth W
KW
D000311:Adrenal Glands / Q000196:embryology / Q000254:growth & development / Q000378:metabolism; D000818:Animals; D004742:Enhancer Elements, Genetic / Q000235:genetics*; D005260:Female; D005455:Fluorescent Antibody Technique; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D007150:Immunohistochemistry; D007668:Kidney / Q000196:embryology / Q000254:growth & development / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D008856:Microscopy, Fluorescence; D009154:Mutation; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics*; D012083:Renin / Q000235:genetics* / Q000378:metabolism; D013363:Submandibular Gland / Q000196:embryology / Q000254:growth & development / Q000378:metabolism; D013997:Time Factors
D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D012083:Renin
Hypertension. 2005 Jan;45(1):3-8
15545507
J Biol Chem. 2005 May 27;280(21):20860-6
15792957
Eur J Biochem. 1997 Jan 15;243(1-2):181-90
9030738
Proc Natl Acad Sci U S A. 1989 Jan;86(1):56-9
2536160
J Virol. 1996 Jul;70(7):4646-54
8676491
Am J Physiol Renal Physiol. 2004 Jun;286(6):F1107-15
14761860
Genesis. 2004 Jan;38(1):39-50
14755803
Kidney Int Suppl. 1998 Sep;67:S12-6
9736246
Physiol Genomics. 2000 Nov 09;4(1):75-81
11074016
J Biol Chem. 2001 Feb 2;276(5):3597-603
11058598
Intern Med. 1999 Apr;38(4):315-23
10361903
Hypertension. 2007 Nov;50(5):933-8
17846348
Circ Res. 2003 May 16;92(9):1033-40
12690040
J Biol Chem. 2007 Oct 12;282(41):29821-30
17690094
J Biol Chem. 2006 Nov 17;281(46):35296-304
16990260
Nucleic Acids Res. 1993 Apr 25;21(8):1687-8
8098522
J Biol Chem. 2004 Jan 9;279(2):1458-67
14563845
Mamm Genome. 2007 Oct;18(10):693-708
17882484
Hypertension. 1991 Oct;18(4):446-57
1916990
Mol Cell Biol. 1984 Nov;4(11):2321-31
6392850
J Biol Chem. 2001 Aug 31;276(35):32489-94
11432851
Circ Res. 1994 May;74(5):764-73
8156625
Circ Res. 1999 Sep 17;85(6):479-88
10488050
Biochim Biophys Acta. 2003 Feb 20;1625(3):280-90
12591615
Nat Biotechnol. 1997 Sep;15(9):859-65
9306400
J Biol Chem. 1996 Sep 13;271(37):22499-505
8798416
J Biol Chem. 2001 Dec 7;276(49):45530-8
11564732
J Biol Chem. 1997 Jan 24;272(4):2412-20
8999953