Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

At a glance: The STEP trials

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.
ADVANCED SEARCH
Log in
Top
Journal of Gastroenterology
Published in: Journal of Gastroenterology 6/2009

01-06-2009 | Original Article—Liver, Pancreas, and Biliary Tract

FIC1-mediated stimulation of FXR activity is decreased with PFIC1 mutations in HepG2 cells

Authors: Saori Koh, Tappei Takada, Ikuya Kukuu, Hiroshi Suzuki

Published in: Journal of Gastroenterology | Issue 6/2009

Login to get access

Abstract

Purpose

Progressive familial intrahepatic cholestasis type 1 (PFIC1) is a specific form of genetic cholestasis caused by functional defects in FIC1/ATP8B1. Although the way FIC1 deficiency leads to PFIC1 remains unclear, some reports suggest that the loss of FIC1 function results in decreased activity of the farnesoid X receptor (FXR) in PFIC1 patients. In this study, in order to elucidate the molecular mechanism of the pathogenesis of PFIC1, we constructed an experimental system for the evaluation of FIC1-mediated stimulatory effects on FXR activity.

Methods and results

Luciferase assays revealed that FIC1 expression increased FXR-dependent transcription and that the effects of three PFIC1 mutants (G308V, T456M and D554N) were smaller than that of wild-type FIC1. In addition, the PFIC1 mutants could not locate to the plasma membrane even in the presence of CDC50A, which brings wild-type FIC1 to the plasma membrane. The results of coprecipitation assays suggested a defect in the ability of the PFIC1 mutants to interact with CDC50A. Furthermore, it was revealed that the expression of CDC50A elevated the FIC1-mediated transcriptional stimulation when coexpressed with wild-type FIC1, but not with mutated FIC1.

Conclusions

These results suggest that the PFIC1 mutants have a lower stimulatory effect on FXR activity and cannot interact with CDC50A, which may lead to the development of the features of PFIC1.
Literature
1.
Muller M, Jansen PL. The secretory function of the liver: new aspects of hepatobiliary transport. J Hepatol. 1998;28:344–54.PubMedCrossRef
2.
Green RM, Hoda F, Ward KL. Molecular cloning and characterization of the murine bile salt export pump. Gene. 2000;241:117–23.PubMedCrossRef
3.
Gerloff T, Stieger B, Hagenbuch B, Madon J, Landmann L, Roth J, et al. The sister of P-glycoprotein represents the canalicular bile salt export pump of mammalian liver. J Biol Chem. 1998;273:10046–50.PubMedCrossRef
4.
Craddock AL, Love MW, Daniel RW, Kirby LC, Walters HC, Wong MH, et al. Expression and transport properties of the human ileal and renal sodium-dependent bile acid transporter. Am J Physiol. 1998;274:G157–69.PubMed
5.
Shneider BL, Dawson PA, Christie DM, Hardikar W, Wong MH, Suchy FJ. Cloning and molecular characterization of the ontogeny of a rat ileal sodium-dependent bile acid transporter. J Clin Invest. 1995;95:745–54.PubMedCrossRef
6.
Wong MH, Oelkers P, Craddock AL, Dawson PA. Expression cloning and characterization of the hamster ileal sodium-dependent bile acid transporter. J Biol Chem. 1994;269:1340–7.PubMed
7.
Dawson PA, Hubbert M, Haywood J, Craddock AL, Zerangue N, Christian WV, et al. The heteromeric organic solute transporter alpha-beta, Ostalpha-Ostbeta, is an ileal basolateral bile acid transporter. J Biol Chem. 2005;280:6960–8.PubMedCrossRef
8.
Rao A, Haywood J, Craddock AL, Belinsky MG, Kruh GD, Dawson PA. The organic solute transporter alpha-beta, Ostalpha-Ostbeta, is essential for intestinal bile acid transport and homeostasis. Proc Natl Acad Sci USA. 2008;105:3891–6.PubMedCrossRef
9.
Hagenbuch B, Meier PJ. Molecular cloning, chromosomal localization, and functional characterization of a human liver Na+/bile acid cotransporter. J Clin Invest. 1994;93:1326–31.PubMedCrossRef
10.
Hagenbuch B, Stieger B, Foguet M, Lubbert H, Meier PJ. Functional expression cloning and characterization of the hepatocyte Na+/bile acid cotransport system. Proc Natl Acad Sci USA. 1991;88:10629–33.PubMedCrossRef
11.
Makishima M, Okamoto AY, Repa JJ, Tu H, Learned RM, Luk A, et al. Identification of a nuclear receptor for bile acids. Science. 1999;284:1362–5.PubMedCrossRef
12.
Parks DJ, Blanchard SG, Bledsoe RK, Chandra G, Consler TG, Kliewer SA, et al. Bile acids: natural ligands for an orphan nuclear receptor. Science. 1999;284:1365–8.PubMedCrossRef
13.
Plass JR, Mol O, Heegsma J, Geuken M, Faber KN, Jansen PL, et al. Farnesoid X receptor and bile salts are involved in transcriptional regulation of the gene encoding the human bile salt export pump. Hepatology. 2002;35:589–96.PubMedCrossRef
14.
Ananthanarayanan M, Balasubramanian N, Makishima M, Mangelsdorf DJ, Suchy FJ. Human bile salt export pump promoter is transactivated by the farnesoid X receptor/bile acid receptor. J Biol Chem. 2001;276:28857–65.PubMedCrossRef
15.
Boyer JL, Trauner M, Mennone A, Soroka CJ, Cai SY, Moustafa T, et al. Upregulation of a basolateral FXR-dependent bile acid efflux transporter OSTalpha-OSTbeta in cholestasis in humans and rodents. Am J Physiol Gastrointest Liver Physiol. 2006;290:G1124–30.PubMedCrossRef
16.
Frankenberg T, Rao A, Chen F, Haywood J, Shneider BL, Dawson PA. Regulation of the mouse organic solute transporter alpha-beta, Ostalpha-Ostbeta, by bile acids. Am J Physiol Gastrointest Liver Physiol. 2006;290:G912–22.PubMedCrossRef
17.
Zollner G, Wagner M, Moustafa T, Fickert P, Silbert D, Gumhold J, et al. Coordinated induction of bile acid detoxification and alternative elimination in mice: role of FXR-regulated organic solute transporter-alpha/beta in the adaptive response to bile acids. Am J Physiol Gastrointest Liver Physiol. 2006;290:G923–32.PubMedCrossRef
18.
Landrier JF, Eloranta JJ, Vavricka SR, Kullak-Ublick GA. The nuclear receptor for bile acids, FXR, transactivates human organic solute transporter-alpha and -beta genes. Am J Physiol Gastrointest Liver Physiol. 2006;290:G476–85.PubMedCrossRef
19.
Lee H, Zhang Y, Lee FY, Nelson SF, Gonzalez FJ, Edwards PA. FXR regulates organic solute transporters alpha and beta in the adrenal gland, kidney, and intestine. J Lipid Res. 2006;47:201–14.PubMedCrossRef
20.
Okuwaki M, Takada T, Iwayanagi Y, Koh S, Kariya Y, Fujii H, et al. LXR alpha transactivates mouse organic solute transporter alpha and beta via IR-1 elements shared with FXR. Pharm Res. 2007;24:390–8.PubMedCrossRef
21.
Chen F, Ma L, Dawson PA, Sinal CJ, Sehayek E, Gonzalez FJ, et al. Liver receptor homologue-1 mediates species- and cell line-specific bile acid-dependent negative feedback regulation of the apical sodium-dependent bile acid transporter. J Biol Chem. 2003;278:19909–16.PubMedCrossRef
22.
Denson LA, Sturm E, Echevarria W, Zimmerman TL, Makishima M, Mangelsdorf DJ, et al. The orphan nuclear receptor, shp, mediates bile acid-induced inhibition of the rat bile acid transporter, ntcp. Gastroenterology. 2001;121:140–7.PubMedCrossRef
23.
Bull LN, van Eijk MJ, Pawlikowska L, DeYoung JA, Juijn JA, Liao M, et al. A gene encoding a P-type ATPase mutated in two forms of hereditary cholestasis. Nat Genet. 1998;18:219–24.PubMedCrossRef
24.
Holthuis JC, Levine TP. Lipid traffic: floppy drives and a superhighway. Nat Rev Mol Cell Biol. 2005;6:209–20.PubMedCrossRef
25.
Paulusma CC, Oude Elferink RP. The type 4 subfamily of P-type ATPases, putative aminophospholipid translocases with a role in human disease. Biochim Biophys Acta. 2005;1741:11–24.PubMed
26.
Chen F, Ananthanarayanan M, Emre S, Neimark E, Bull LN, Knisely AS, et al. Progressive familial intrahepatic cholestasis, type 1, is associated with decreased farnesoid X receptor activity. Gastroenterology. 2004;126:756–64.PubMedCrossRef
27.
Alvarez L, Jara P, Sanchez-Sabate E, Hierro L, Larrauri J, Diaz MC, et al. Reduced hepatic expression of farnesoid X receptor in hereditary cholestasis associated to mutation in ATP8B1. Hum Mol Genet. 2004;13:2451–60.PubMedCrossRef
28.
Noji T, Yamamoto T, Saito K, Fujimura-Kamada K, Kondo S, Tanaka K. Mutational analysis of the Lem3p-Dnf1p putative phospholipid-translocating P-type ATPase reveals novel regulatory roles for Lem3p and a carboxyl-terminal region of Dnf1p independent of the phospholipid-translocating activity of Dnf1p in yeast. Biochem Biophys Res Commun. 2006;344:323–31.PubMedCrossRef
29.
Chen S, Wang J, Muthusamy BP, Liu K, Zare S, Andersen RJ, et al. Roles for the Drs2p-Cdc50p complex in protein transport and phosphatidylserine asymmetry of the yeast plasma membrane. Traffic. 2006;7:1503–17.PubMedCrossRef
30.
Saito K, Fujimura-Kamada K, Furuta N, Kato U, Umeda M, Tanaka K. Cdc50p, a protein required for polarized growth, associates with the Drs2p P-type ATPase implicated in phospholipid translocation in Saccharomyces cerevisiae. Mol Biol Cell. 2004;15:3418–32.PubMedCrossRef
31.
Paulusma CC, Folmer DE, Ho-Mok KS, de Waart DR, Hilarius PM, Verhoeven AJ, et al. ATP8B1 requires an accessory protein for endoplasmic reticulum exit and plasma membrane lipid flippase activity. Hepatology. 2008;47:268–78.PubMedCrossRef
32.
Noe J, Stieger B, Meier PJ. Functional expression of the canalicular bile salt export pump of human liver. Gastroenterology. 2002;123:1659–66.PubMedCrossRef
33.
Hayashi H, Takada T, Suzuki H, Akita H, Sugiyama Y. Two common PFIC2 mutations are associated with the impaired membrane trafficking of BSEP/ABCB11. Hepatology. 2005;41:916–24.PubMedCrossRef
34.
Iwayanagi Y, Takada T, Suzuki H. HNF4alpha is a crucial modulator of the cholesterol-dependent regulation of NPC1L1. Pharm Res. 2008;25:1134–41.PubMedCrossRef
35.
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951;193:265–75.PubMed
36.
Klomp LW, Vargas JC, van Mil SW, Pawlikowska L, Strautnieks SS, van Eijk MJ, et al. Characterization of mutations in ATP8B1 associated with hereditary cholestasis. Hepatology. 2004;40:27–38.PubMedCrossRef
37.
van Mil SW, van Oort MM, van den Berg IE, Berger R, Houwen RH, Klomp LW. Fic1 is expressed at apical membranes of different epithelial cells in the digestive tract and is induced in the small intestine during postnatal development of mice. Pediatr Res. 2004;56:981–7.PubMedCrossRef
38.
Eppens EF, van Mil SW, de Vree JM, Mok KS, Juijn JA, Oude Elferink RP, et al. FIC1, the protein affected in two forms of hereditary cholestasis, is localized in the cholangiocyte and the canalicular membrane of the hepatocyte. J Hepatol. 2001;35:436–43.PubMedCrossRef
39.
Ujhazy P, Ortiz D, Misra S, Li S, Moseley J, Jones H, et al. Familial intrahepatic cholestasis 1: studies of localization and function. Hepatology. 2001;34:768–75.PubMedCrossRef
40.
Pawlikowska L, Groen A, Eppens EF, Kunne C, Ottenhoff R, Looije N, et al. A mouse genetic model for familial cholestasis caused by ATP8B1 mutations reveals perturbed bile salt homeostasis but no impairment in bile secretion. Hum Mol Genet. 2004;13:881–92.PubMedCrossRef
41.
Paulusma CC, Groen A, Kunne C, Ho-Mok KS, Spijkerboer AL, Rudi de Waart D, et al. Atp8b1 deficiency in mice reduces resistance of the canalicular membrane to hydrophobic bile salts and impairs bile salt transport. Hepatology. 2006;44:195–204.PubMedCrossRef
42.
Lykavieris P, van Mil S, Cresteil D, Fabre M, Hadchouel M, Klomp L, et al. Progressive familial intrahepatic cholestasis type 1 and extrahepatic features: no catch-up of stature growth, exacerbation of diarrhea, and appearance of liver steatosis after liver transplantation. J Hepatol. 2003;39:447–52.PubMedCrossRef
43.
Knisely AS. Progressive familial intrahepatic cholestasis: a personal perspective. Pediatr Dev Pathol. 2000;3:113–25.PubMedCrossRef
44.
Groen A, Kunne C, Paulusma CC, Kramer W, Agellon LB, Bull LN, et al. Intestinal bile salt absorption in Atp8b1 deficient mice. J Hepatol. 2007;47:114–22.PubMedCrossRef
45.
Gineste R, Sirvent A, Paumelle R, Helleboid S, Aquilina A, Darteil R, et al. Phosphorylation of farnesoid X receptor by protein kinase C promotes its transcriptional activity. Mol Endocrinol. 2008;22:2433–47.PubMedCrossRef
46.
Frankenberg T, Miloh T, Chen FY, Ananthanarayanan M, Sun AQ, Balasubramaniyan N, et al. The membrane protein ATPase class I type 8B member 1 signals through protein kinase C zeta to activate the farnesoid X receptor. Hepatology. 2008;48:1896–905.PubMedCrossRef
Metadata
Title
FIC1-mediated stimulation of FXR activity is decreased with PFIC1 mutations in HepG2 cells
Authors
Saori Koh
Tappei Takada
Ikuya Kukuu
Hiroshi Suzuki
Publication date
01-06-2009
Publisher
Springer Japan
Published in
Journal of Gastroenterology / Issue 6/2009
Print ISSN: 0944-1174
Electronic ISSN: 1435-5922
DOI
https://doi.org/10.1007/s00535-009-0041-y

Other articles of this Issue 6/2009

Journal of Gastroenterology 6/2009 Go to the issue

Original Article—Alimentary Tract

Sargramostim in patients with Crohn’s disease: results of a phase 1–2 study

Original Article—Alimentary Tract

Down-regulation of miR-141 in gastric cancer and its involvement in cell growth

Original Article—Liver, Pancreas, and Biliary Tract

Preoperative serum gamma-glutamyl transferase to alanine aminotransferase ratio is a convenient prognostic marker for Child-Pugh A hepatocellular carcinoma after operation

Original Article—Liver, Pancreas, and Biliary Tract

Efficacy and safety of addition of minor bloodletting (petit phlebotomy) in hepatitis C virus-infected patients receiving regular glycyrrhizin injections

Original Article—Alimentary Tract

Prevalence of mid-gastrointestinal bleeding in patients with acute overt gastrointestinal bleeding: multi-center experience with 1,044 consecutive patients

Original Article—Alimentary Tract

Usefulness of endoscopic ultrasound-guided fine needle aspiration (EUS-FNA) for undiagnosed intra-abdominal lymphadenopathy
Obesity Clinical Trial Summary

At a glance: The STEP trials

Read more

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.

Developed by: Springer Medicine

Highlights from the ACC 2024 Congress

Year in Review: Pediatric cardiology

Pediatric Cardiology Video

Watch Dr. Anne Marie Valente present the last year's highlights in pediatric and congenital heart disease in the official ACC.24 Year in Review session.

Year in Review: Pulmonary vascular disease

Cardiopulmonary Comorbidity Video

The last year's highlights in pulmonary vascular disease are presented by Dr. Jane Leopold in this official video from ACC.24.

Year in Review: Valvular heart disease

Valvular Heart Disease Video

Watch Prof. William Zoghbi present the last year's highlights in valvular heart disease from the official ACC.24 Year in Review session.

Year in Review: Heart failure and cardiomyopathies

Heart Failure Video

Watch this official video from ACC.24. Dr. Biykem Bozkurt discuss last year's major advances in heart failure and cardiomyopathies.

ACC 2024 Congress Coverage

Heart Failure Video

Year in Review: Heart failure and cardiomyopathies

Watch now

Watch this official video from ACC.24. Dr. Biykem Bozkurt discuss last year's major advances in heart failure and cardiomyopathies.

Semaglutide benefits people with type 2 diabetes and obesity-related heart failure

16-04-2024 Type 2 Diabetes News

Incentivised to exercise

11-04-2024 Lifestyle Modifications News

New intervention for STEMI-related cardiogenic shock

10-04-2024 Myocardial Infarction News

» View more highlights on the ACC 2024 congress hub page

Image Credits