title: Mutation of the palmitoylation site of estrogen receptor α in vivo reveals tissue-specific roles for membrane versus nuclear actions.
abstract: Estrogen receptor alpha (ERα) activation functions AF-1 and AF-2 classically mediate gene transcription in response to estradiol (E2). A fraction of ERα is targeted to plasma membrane and elicits membrane-initiated steroid signaling (MISS), but the physiological roles of MISS in vivo are poorly understood. We therefore generated a mouse with a point mutation of the palmitoylation site of ERα (C451A-ERα) to obtain membrane-specific loss of function of ERα. The abrogation of membrane localization of ERα in vivo was confirmed in primary hepatocytes, and it resulted in female infertility with abnormal ovaries lacking corpora lutea and increase in luteinizing hormone levels. In contrast, E2 action in the uterus was preserved in C451A-ERα mice and endometrial epithelial proliferation was similar to wild type. However, E2 vascular actions such as rapid dilatation, acceleration of endothelial repair, and endothelial NO synthase phosphorylation were abrogated in C451A-ERα mice. A complementary mutant mouse lacking the transactivation function AF-2 of ERα (ERα-AF2(0)) provided selective loss of function of nuclear ERα actions. In ERα-AF2(0), the acceleration of endothelial repair in response to estrogen-dendrimer conjugate, which is a membrane-selective ER ligand, was unaltered, demonstrating integrity of MISS actions. In genome-wide analysis of uterine gene expression, the vast majority of E2-dependent gene regulation was abrogated in ERα-AF2(0), whereas in C451A-ERα it was nearly fully preserved, indicating that membrane-to-nuclear receptor cross-talk in vivo is modest in the uterus. Thus, this work genetically segregated membrane versus nuclear actions of a steroid hormone receptor and demonstrated their in vivo tissue-specific roles.
authors.affiliation: Institut National de la Santé et de la Recherche Médicale U1048, Institut des Maladies Métaboliques et Cardiovasculaires, Université de Toulouse, 31 432 Toulouse Cedex 4, France.
authors.forename: Marine
authors.initials: M
authors.lastname: Adlanmerini
authors.forename: Romain
authors.initials: R
authors.lastname: Solinhac
authors.forename: Anne
authors.initials: A
authors.lastname: Abot
authors.forename: Aurélie
authors.initials: A
authors.lastname: Fabre
authors.forename: Isabelle
authors.initials: I
authors.lastname: Raymond-Letron
authors.forename: Anne-Laure
authors.initials: AL
authors.lastname: Guihot
authors.forename: Frédéric
authors.initials: F
authors.lastname: Boudou
authors.forename: Lucile
authors.initials: L
authors.lastname: Sautier
authors.forename: Emilie
authors.initials: E
authors.lastname: Vessières
authors.forename: Sung Hoon
authors.initials: SH
authors.lastname: Kim
authors.forename: Philippe
authors.initials: P
authors.lastname: Lière
authors.forename: Coralie
authors.initials: C
authors.lastname: Fontaine
authors.forename: Andrée
authors.initials: A
authors.lastname: Krust
authors.forename: Pierre
authors.initials: P
authors.lastname: Chambon
authors.forename: John A
authors.initials: JA
authors.lastname: Katzenellenbogen
authors.forename: Pierre
authors.initials: P
authors.lastname: Gourdy
authors.forename: Philip W
authors.initials: PW
authors.lastname: Shaul
authors.forename: Daniel
authors.initials: D
authors.lastname: Henrion
authors.forename: Jean-François
authors.initials: JF
authors.lastname: Arnal
authors.forename: Françoise
authors.initials: F
authors.lastname: Lenfant
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D015153:Blotting, Western; D002462:Cell Membrane / Q000378:metabolism*; D002465:Cell Movement; D002467:Cell Nucleus / Q000378:metabolism*; D019295:Computational Biology; D042783:Endothelial Cells; D047628:Estrogen Receptor alpha / Q000235:genetics* / Q000378:metabolism; D005260:Female; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry; D054878:Lipoylation / Q000235:genetics; D051379:Mice; D008822:Mice, Transgenic; D046228:Microarray Analysis; D010053:Ovary / Q000378:metabolism / Q000502:physiology*; D017354:Point Mutation / Q000235:genetics; D020239:Receptor Cross-Talk / Q000502:physiology; D014599:Uterus / Q000378:metabolism*
chemical_list: D047628:Estrogen Receptor alpha
keywords: fertility; genomic actions; nongenomic effects; vascular effects