Emery-Dreifuss muscular dystrophy mutations impair TRC40-mediated targeting of emerin to the inner nuclear membrane
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Emery-Dreifuss muscular dystrophy mutations impair TRC40-mediated targeting of emerin to the inner nuclear membrane. / Pfaff, Janine; Rivera Monroy, Jhon; Jamieson, Cara; Rajanala, Kalpana; Vilardi, Fabio; Schwappach, Blanche; Kehlenbach, Ralph H.
in: J CELL SCI, Jahrgang 129, Nr. 3, 01.02.2016, S. 502-16.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
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T1 - Emery-Dreifuss muscular dystrophy mutations impair TRC40-mediated targeting of emerin to the inner nuclear membrane
AU - Pfaff, Janine
AU - Rivera Monroy, Jhon
AU - Jamieson, Cara
AU - Rajanala, Kalpana
AU - Vilardi, Fabio
AU - Schwappach, Blanche
AU - Kehlenbach, Ralph H
N1 - © 2016. Published by The Company of Biologists Ltd.
PY - 2016/2/1
Y1 - 2016/2/1
N2 - Emerin is a tail-anchored protein that is found predominantly at the inner nuclear membrane (INM), where it associates with components of the nuclear lamina. Mutations in the emerin gene cause Emery-Dreifuss muscular dystrophy (EDMD), an X-linked recessive disease. Here, we report that the TRC40/GET pathway for post-translational insertion of tail-anchored proteins into membranes is involved in emerin-trafficking. Using proximity ligation assays, we show that emerin interacts with TRC40 in situ. Emerin expressed in bacteria or in a cell-free lysate was inserted into microsomal membranes in an ATP- and TRC40-dependent manner. Dominant-negative fragments of the TRC40-receptor proteins WRB and CAML (also known as CAMLG) inhibited membrane insertion. A rapamycin-based dimerization assay revealed correct transport of wild-type emerin to the INM, whereas TRC40-binding, membrane integration and INM-targeting of emerin mutant proteins that occur in EDMD was disturbed. Our results suggest that the mode of membrane integration contributes to correct targeting of emerin to the INM.
AB - Emerin is a tail-anchored protein that is found predominantly at the inner nuclear membrane (INM), where it associates with components of the nuclear lamina. Mutations in the emerin gene cause Emery-Dreifuss muscular dystrophy (EDMD), an X-linked recessive disease. Here, we report that the TRC40/GET pathway for post-translational insertion of tail-anchored proteins into membranes is involved in emerin-trafficking. Using proximity ligation assays, we show that emerin interacts with TRC40 in situ. Emerin expressed in bacteria or in a cell-free lysate was inserted into microsomal membranes in an ATP- and TRC40-dependent manner. Dominant-negative fragments of the TRC40-receptor proteins WRB and CAML (also known as CAMLG) inhibited membrane insertion. A rapamycin-based dimerization assay revealed correct transport of wild-type emerin to the INM, whereas TRC40-binding, membrane integration and INM-targeting of emerin mutant proteins that occur in EDMD was disturbed. Our results suggest that the mode of membrane integration contributes to correct targeting of emerin to the INM.
KW - Adenosine Triphosphate/metabolism
KW - Arsenite Transporting ATPases/metabolism
KW - Cell Line, Tumor
KW - HeLa Cells
KW - Humans
KW - Membrane Proteins/metabolism
KW - Microsomes/metabolism
KW - Muscular Dystrophy, Emery-Dreifuss/genetics
KW - Mutation/genetics
KW - Nuclear Envelope/metabolism
KW - Nuclear Proteins/metabolism
KW - Protein Binding/genetics
KW - Protein Processing, Post-Translational/genetics
KW - Protein Transport/genetics
U2 - 10.1242/jcs.179333
DO - 10.1242/jcs.179333
M3 - SCORING: Journal article
C2 - 26675233
VL - 129
SP - 502
EP - 516
JO - J CELL SCI
JF - J CELL SCI
SN - 0021-9533
IS - 3
ER -
