title: Differential effects of progressive familial intrahepatic cholestasis type 1 and benign recurrent intrahepatic cholestasis type 1 mutations on canalicular localization of ATP8B1.
abstract: Mutations in ATP8B1 cause progressive familial intrahepatic cholestasis type 1 (PFIC1) and benign recurrent intrahepatic cholestasis type 1 (BRIC1), forming a spectrum of cholestatic disease. Whereas PFIC1 is a progressive, endstage liver disease, BRIC1 patients suffer from episodic periods of cholestasis that resolve spontaneously. At present it is not clear how the type and location of the mutations relate to the clinical manifestations of PFIC1 and BRIC1. ATP8B1 localizes to the canalicular membrane of hepatocytes where it mediates the inward translocation of phosphatidylserine. ATP8B1 interacts with CDC50A, which is required for endoplasmic reticulum exit and plasma membrane localization. In this study we analyzed a panel of missense mutations causing PFIC1 (G308V, D554N, G1040R) or BRIC1 (D70N, I661T). In addition, we included two mutations that have been associated with intrahepatic cholestasis of pregnancy (ICP) (D70N, R867C). We examined the effect of these mutations on protein stability and interaction with CDC50A in Chinese hamster ovary cells, and studied the subcellular localization in WIF-B9 cells. Protein stability was reduced for three out of six mutations studied. Two out of three PFIC1 mutant proteins did not interact with CDC50A, whereas BRIC1/ICP mutants displayed reduced interaction. Importantly, none of the PFIC1 mutants were detectable in the canalicular membrane of WIF-B9 cells, whereas all BRIC1/ICP mutants displayed the same cellular staining pattern as wild-type ATP8B1. Our data indicate that PFIC1 mutations lead to the complete absence of canalicular expression, whereas in BRIC1/ICP residual protein is expressed in the canalicular membrane.
  
  
  CONCLUSIONS
  These data provide an explanation for the difference in severity between the phenotypes of PFIC1 and BRIC1.
authors.affiliation: AMC Liver Center, Academic Medical Center, Amsterdam, The Netherlands.
authors.forename: Dineke E
authors.initials: DE
authors.lastname: Folmer
authors.forename: Vincent A
authors.initials: VA
authors.lastname: van der Mark
authors.forename: Kam S
authors.initials: KS
authors.lastname: Ho-Mok
authors.forename: Ronald P J
authors.initials: RP
authors.lastname: Oude Elferink
authors.forename: Coen C
authors.initials: CC
authors.lastname: Paulusma
mesh_terms: D000251:Adenosine Triphosphatases / Q000235:genetics* / Q000378:metabolism*; D000818:Animals; D002460:Cell Line; D002780:Cholestasis, Intrahepatic / Q000378:metabolism / Q000473:pathology; D006224:Cricetinae; D003412:Cricetulus; D004195:Disease Models, Animal; D005260:Female; D022781:Hepatocytes / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism; D020125:Mutation, Missense / Q000235:genetics*; D049294:Phospholipid Transfer Proteins; D051381:Rats
chemical_list: C539644:Atp8b1 protein, rat; D008565:Membrane Proteins; D049294:Phospholipid Transfer Proteins; C526978:TMEM30a protein, human; C545244:Tmem30a protein, rat; D000251:Adenosine Triphosphatases; C111067:ATP8B1 protein, human; C545243:PFIC1 protein, rat