The CAAT-binding transcription factor 1/nuclear factor 1 binding site is important in beta-myosin heavy chain antisense promoter regulation in rats.
The rat heart expresses two myosin heavy chain (MHC) isoforms, beta and alpha; these genes are arranged in tandem on the same chromosome. We have reported that an antisense (AS) beta RNA starts in the intergenic (IG) region between beta and alpha genes and extends to overlap the beta gene. We propose that in adult rats, both the alpha sense and IG betaAS RNA expression are activated by an IG bidirectional promoter and that the transcription of betaAS RNA interferes with the sense beta, resulting in low levels of beta mRNA and high levels of alpha, a phenotype seen in a typical rat heart. A previous report examined the activity of the betaAS promoter and showed that a 559 bp fragment of the betaAS promoter (-2285 to -1726; relative to alphaMHC gene start site) injected into rat ventricle was activated in control heart, and decreased significantly in response to hypothyroidism (propylthiouracil induced) and diabetes (streptozotocin induced) and increased in hyperthyroid rats (T(3) induced), similar in pattern to the endogenous betaAS RNA. In the present paper, we demonstrate with electrophoretic mobility shift analyses that ventricular nuclear proteins are interacting with a nuclear factor 1/CAAT-binding transcription factor 1 (NF1/CTF1) binding site, and a supershift assay indicates that the protein binding at this site is antigenetically related to the CTF1/NF1 factor. Moreover, a mutation of the CTF1/NF1 site within the 559 bp promoter region nearly abolished promoter activity in vivo in control, STZ- and PTU-treated rats. Based on these findings, we conclude that the NF1 site is critical to betaAS promoter regulation.
Giger
Julia M
JM
Department of Physiology and Biophysics, University of California, Irvine, D-346, Medical Sciences Building I, Irvine, CA 92697, USA. jmeehan@uci.edu
Bodell
Paul W
PW
Baldwin
Kenneth M
KM
Haddad
Fadia
F
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D003921:Diabetes Mellitus, Experimental / Q000503:physiopathology; D005260:Female; D006980:Hyperthyroidism / Q000503:physiopathology; D007037:Hypothyroidism / Q000503:physiopathology; D008969:Molecular Sequence Data; D018995:Myosin Heavy Chains / Q000235:genetics*; D051839:NFI Transcription Factors / Q000502:physiology*; D011401:Promoter Regions, Genetic; D016372:RNA, Antisense / Q000502:physiology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013045:Species Specificity; D013963:Thyroid Hormones / Q000502:physiology; D024723:Ventricular Myosins / Q000235:genetics*
D051839:NFI Transcription Factors; D016372:RNA, Antisense; D013963:Thyroid Hormones; C080333:transcription factor nuclear factor 1; D024723:Ventricular Myosins; D018995:Myosin Heavy Chains
J Biol Chem. 1999 Apr 30;274(18):12917-24
10212282
FEBS Lett. 1979 Jan 15;97(2):330-4
761639
J Biol Chem. 2003 Sep 26;278(39):37132-8
12851393
J Clin Invest. 1989 Dec;84(6):1693-700
2687327
J Clin Invest. 1997 Nov 1;100(9):2315-24
9410910
Gene Expr. 1993;3(1):11-25
8508026
Microsc Res Tech. 2000 Sep 15;50(6):522-31
10998641
Physiol Rev. 1996 Apr;76(2):371-423
8618961
Biotechniques. 1999 Feb;26(2):202-4, 206
10023525
Circ Res. 1995 Aug;77(2):439-44
7614728
J Biol Chem. 1993 Feb 25;268(6):4331-6
8440718
J Biol Chem. 1984 May 25;259(10):6437-46
6327679
Am J Physiol Endocrinol Metab. 2000 Dec;279(6):E1319-24
11093920
Nature. 1988 Jul 21;334(6179):218-24
3398920
J Clin Invest. 1997 Nov 1;100(9):2362-70
9410916
Endocrinology. 1996 Mar;137(3):802-8
8603588
Mol Cell Biol. 2004 Apr;24(7):3036-47
15024090
Circ Res. 1991 Mar;68(3):836-46
1742869
J Biol Chem. 1999 May 21;274(21):15213-21
10329730
J Mol Cell Cardiol. 1984 Mar;16(3):203-18
6232394
J Appl Physiol (1985). 2005 Apr;98(4):1185-94
15591287
J Mol Biol. 1995 Oct 13;253(1):17-31
7473711
Am J Physiol. 1995 Oct;269(4 Pt 1):C1025-33
7485442
Biochem Biophys Res Commun. 1991 Sep 30;179(3):1269-75
1930172
Am J Physiol Heart Circ Physiol. 2007 Jun;292(6):H3065-71
17307996
N Engl J Med. 2002 May 2;346(18):1357-65
11986409
Biochem Biophys Res Commun. 1999 Jul 14;260(3):600-4
10403812
DNA Seq. 2005 Apr;16(2):151-5
16147868
Differentiation. 2002 Oct;70(8):457-67
12366383
Biochem Biophys Res Commun. 1999 Nov;265(1):222-32
10548518
Nature. 1982 Jun 24;297(5868):659-64
7045682
Circ Res. 1982 Apr;50(4):491-500
6461437
Am J Physiol Heart Circ Physiol. 2004 Jul;287(1):H408-13
15001437
Proc Natl Acad Sci U S A. 1984 May;81(9):2626-30
6585819
Am J Physiol Heart Circ Physiol. 2008 Jan;294(1):H29-40
17982008
J Biol Chem. 1996 Jun 21;271(25):15008-17
8663099