title: The CAAT-binding transcription factor 1/nuclear factor 1 binding site is important in beta-myosin heavy chain antisense promoter regulation in rats.
abstract: 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.
authors.affiliation: Department of Physiology and Biophysics, University of California, Irvine, D-346, Medical Sciences Building I, Irvine, CA 92697, USA. jmeehan@uci.edu
authors.forename: Julia M
authors.initials: JM
authors.lastname: Giger
authors.forename: Paul W
authors.initials: PW
authors.lastname: Bodell
authors.forename: Kenneth M
authors.initials: KM
authors.lastname: Baldwin
authors.forename: Fadia
authors.initials: F
authors.lastname: Haddad
mesh_terms: 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*
chemical_list: D051839:NFI Transcription Factors; D016372:RNA, Antisense; D013963:Thyroid Hormones; C080333:transcription factor nuclear factor 1; D024723:Ventricular Myosins; D018995:Myosin Heavy Chains
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