Cloning and sequencing of cDNAs encoding the human hepatocyte nuclear factor 4 indicate the presence of two isoforms in human liver

doi:10.1016/0378-1119(94)90079-5

Gene

Volume 147, Issue 2, 30 September 1994, Pages 269-272

https://doi.org/10.1016/0378-1119(94)90079-5 Get rights and content

Abstract

Hepatocyte nuclear factor 4 (HNF-4) is a key transcription factor involved in the specific expression of many genes in liver and intestine. Sequences of cDNAs coding for HNF-4 have been established in rat and Drosophila melanogaster. Rat HNF-4 exhibits two isoforms which probably result from differential splicing. We have isolated HNF-4 cDNAs from an adult human cDNA library. Sequence analysis revealed that two HNF-4 isoforms are also present in human liver. The complete sequence of the longest human isoform has been established and compared to the rat HNF-4 amino-acid sequences.

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Cited by (79)

The interplay between hepatocyte nuclear factor 4α (HNF4α) and cholesterol sulfotransferase (SULT2B1b) in hepatic energy homeostasis
2019, Liver Research
Citation Excerpt :
Through their regulation of the expression of target genes, nuclear receptors can regulate a wide array of physiological processes, including energy metabolism.1 HNF4α, which is also known as NR2A1 (nuclear receptor subfamily 2, group A, member 1), is a nuclear receptor encoded by the HNF4A gene.2,3 HNF4α binds to deoxyribonucleic acid (DNA) as a homodimer.1
The nuclear receptor hepatocyte nuclear factor 4alpha (HNF4α) plays a critical role in the regulation of metabolic homeostasis, including glucose homeostasis. Sulfotransferases (SULTs) catalyze the transfer of a sulfate group from 3-phosphoadenosine 5-phosphosulfate (PAPS) to an acceptor molecule. Sulfonation plays an essential role in regulating the chemical and functional homeostasis of endogenous and exogenous molecules. Among SULTs, the cholesterol sulfotransferase 2B1b (SULT2B1b) preferentially catalyzes the sulfoconjugation of cholesterol and oxysterols to form cholesterol sulfate and oxysterol sulfates. Hepatic gluconeogenesis represents a critical component of energy metabolism. Although there have been reviews on the regulation of glucose homeostasis by HNF4α, the interplay between HNF4α and SULT2B1b in hepatic glucose homeostasis remains scattered. In this review, we intend to provide an overview on how HNF4α functionally cross-talks with SULT2B1b to regulate hepatic glucose homeostasis and whether the HNF4α-SULT2B1b axis represents a novel therapeutic target for the management of metabolic liver disease and metabolic syndrome.
Congenital hyperinsulinism and glycogenosis-like phenotype due to a novel HNF4A mutation
2017, Diabetes Research and Clinical Practice
Citation Excerpt :
On the other hand, hepatomegaly and increased liver transaminase levels of our proband are not common in HNF4A mutation carriers. However, these features are in accordance with results in liver-specific HNF4A knock-out mice [19], and also with several previously reported patients with CHI carrying the p.R63W mutation (originally published as p.R76W, numbering according to Chartier et al. [20]) in the HNF4A gene [6,21,22] (Table 2). As the combination of CHI and hepatopathy is rare, such patients could benefit from the genetic analysis of HNF4A gene.
Congenital hyperinsulinism (CHI) and glycogen storage disease (glycogenosis) are both causing hypoglycemia during infancy, but with different additional clinical features and therapeutic approach. We aimed to identify a genetic cause in a child with an ambiguous phenotype.
We present a child with hyperinsulinemic hypoglycemia, physiological 3-OH butyrate, increased triglyceride serum levels, increased level of glycogen in erythrocytes, increased liver transaminases, and increased echogenicity on liver ultrasonography. As both parents of the proband were referred as healthy, we raised a clinical suspicion on glycogenosis with recessive inheritance. However, whole exome sequencing revealed no mutation in genes causing glycogenosis, but a novel heterozygous variant LRG_483t1: c.427-1G > A in the HNF4A gene was identified. Aberrant splicing resulting in in-frame deletion c.429_476del, p.(T144_I159del) was confirmed by sequencing of HNF4A transcripts reverse-transcribed from whole blood RNA. The same variant was found in five of eight tested family relatives (one of them already had diabetes, two had prediabetes). With regard to the results of DNA analysis, we added diazoxide to the therapy. Consequently, the frequency and severity of hypoglycemia in the proband decreased. We have also recommended sulfonylurea treatment after diabetes onset in adult mutation carriers.
We have identified a novel HNF4A gene mutation in our patient with CHI and glycogenosis-like phenotype. The proband and her family members benefited from the genetic testing by WES method and consequently personalized therapy. Nevertheless, the HNF4A gene testing may be considered in selected CHI cases with glycogenosis-like phenotype prior WES analysis.
Two patients with HNF4A-related congenital hyperinsulinism and renal tubular dysfunction: A clinical variation which includes transient hepatic dysfunction
2015, Diabetes Research and Clinical Practice
Citation Excerpt :
After informed consent was provided from the guardians, HNF4A mutational analysis was performed as described previously [3]. Both cases carried a heterozygous p.R76W mutation in HNF4A according to conventional cDNA numbering [4]. Their parents did not have the mutation.
Hepatocyte nuclear factor 4α regulates the expression of the murine pyruvate carboxylase gene through the HNF4-specific binding motif in its proximal promoter
2013, Biochimica et Biophysica Acta - Gene Regulatory Mechanisms
Citation Excerpt :
Total RNA was isolated from 1 × 106 HepG2 cells using Trizol reagent (Invitrogen). cDNA synthesis was performed in a 20 μl reaction mixture containing 2 μg of heat denatured total RNA, 200 ng random hexamer, 1 mM dNTP, 1 × reverse transcriptase buffer (50 mM Tris pH 8.3, 75 mM KCl, 1 mM DTT) and 200 units of SuperscriptIII reverse transcriptase (Invitrogen) at 50 °C for 1 h. hHNF4α cDNA was cloned by PCR using forward and reverse primers designed from the published sequence [18]. PCR was carried out in a 50 μl reaction mixture containing 1 × PCR buffer (20 mM Tris, pH 8.4, 50 mM MgCl2, 1.5 mM, MgCl2), 0.2 mM dNTP, 5 ng cDNA, 0.5 μM HNF-F (5′-AAGCTTATGCGACTCTCCAAAACCTCG-3′, underline indicates HindIII restriction site) and HNF-R (5′-GGTACCCTAGATAACTTCCTGCTTGTG-3′, underline indicates KpnI restriction site) primers, and 2.5 units of Taq DNA polymerase (Invitrogen).
Pyruvate carboxylase (PC) is the first regulatory enzyme of gluconeogenesis. Here we report that the proximal promoter of the murine PC gene contains three binding sites for hepatocyte nuclear factor 4α (HNF4α). These sites include the classical direct repeat 1 (DR1) (− 386/− 374), non-perfect DR1 (− 118/− 106) and HNF4α-specific binding motif (H4-SBM) (− 26/− 14). Under basal conditions, mutation of the non-perfect DR1 decreased promoter activity by 50%, whereas mutation of neither the DR1 nor the H4-SBM had any effect. In marked contrast, only mutation of the H4-SBM decreased HNF4α-transactivation of the promoter activity by 65%. EMSA revealed that HNF4α binds to the DR1site and H4-SBM with similar affinity while it binds poorly to the non-perfect DR1. Interestingly, this non-perfect DR1 also coincides with two E-boxes. Mutation of the non-perfect DR1 together with the nearby E-box reduced USF1- but not USF2-transactivation of promoter activity, suggesting that USF1 partly contributes to the basal activity of the promoter. Substitution of the H4-SBM with the DR1 marginally reduced the basal promoter activity but did not eliminate HNF4α-transactivation, suggesting that HNF4α can exert its effect via DR1 within this promoter context. ChIP-assay confirmed that HNF4α is associated with the H4-SBM. Suppression of HNF4α expression in AML12 cells down-regulated PC mRNA and PC protein by 60% and 50%, respectively, confirming that PC is a target of HNF4α. We also propose a model for differential regulation of P1 promoter of PC gene in adipose tissue and liver.
Hepatocyte nuclear factor 4α multiple isoforms, their functions, and their interactomes
2023, Proteomics
Transcription factor HNF4α2 promotes osteogenesis and prevents bone abnormalities in mice with renal osteodystrophy
2023, Journal of Clinical Investigation

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Cloning and sequencing of cDNAs encoding the human hepatocyte nuclear factor 4 indicate the presence of two isoforms in human liver

Abstract

Anal. Biochem.

Biochim. Biophys. Acta

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

More potent transcriptional activators or a transdominant inhibitor of the HNF1 homeoprotein family are generated by alternative RNA processing

EMBO J.

Regulated expression of three C/EBP isoforms during adipose conversion of 3T3-L1 cells

Genes Dev.

HNF-4 increases activity of the rat Apo A1 gene

Nucleic Acids Res.

Compilation of vertebrate-encoded transcription factors

Nucleic Acids Res.

Reciprocal regulation of adipogenesis by Myc and C/EBP alpha

Science