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Ultraspiracle promotes the nuclear localization of ecdysteroid receptor in mammalian cells

Ultraspiracle promotes the nuclear localization of ecdysteroid receptor in mammalian cells
Ultraspiracle promotes the nuclear localization of ecdysteroid receptor in mammalian cells
The heterodimer consisting of ecdysteroid receptor (EcR) and ultraspiracle (USP), both of which are members of the nuclear receptor superfamily, is considered to be the functional ecdysteroid receptor. Here we analyzed the subcellular distribution of EcR and USP fused to fluorescent proteins. The experiments were carried out in mammalian COS-7, CHO-K1 and HeLa cells to facilitate investigation of the subcellular trafficking of EcR and USP in the absence of endogenous expression of these two receptors. The distribution of USP tagged with a yellow fluorescent protein (YFP-USP) was almost exclusively nuclear in all cell types analyzed. The nuclear localization remained constant for at least 1 day after the first visible signs of expression. In contrast, the intracellular distribution of EcR tagged with a yellow fluorescent protein (YFP-EcR) varied and was dependent on time and cell type, although YFP-EcR alone was also able to partially translocate into the nuclear compartment. Coexpression of YFP-EcR with USP tagged with a cyan fluorescent protein (CFP-USP) resulted in exclusively nuclear localization of both proteins in all cell types analyzed. The USP-induced nuclear localization of YFP-EcR was stable for at least 20 hours. These experiments suggest that USP has a profound effect on the subcellular distribution of EcR.
ecdysone, green fluorescent protein, nuclear receptor, subcellular distribution
1431-6730
463-470
Nieva, C.
a8fc81a0-512a-4bea-863d-c896cf37a2a9
Gwoźdź, T.
ce8ac030-3ae8-4191-bd27-36a2c458ab87
Dutko-Gwoźdź, J.
a3f2ee57-5f8c-46e5-88f0-f9ef16cc9667
Wiedenmann, J.
ad445af2-680f-4927-90b3-589ac9d538f7
Spindler-Barth, M.
1bcdd718-ca1a-4e79-8193-954b6150bd2d
Wieczorek, E.
172d17d7-a902-463c-b6c2-720d26c69537
Dobrucki, J.
615b2dbd-a401-4ae0-9ee4-1114edc47ed6
Duś, D.
44b73782-7c26-4266-a594-e03578dc2313
Henrich, V.
524e2151-9aa4-4004-a474-823d392776db
Ożyhar, A.
f60f3cb9-7823-42c2-8da6-7db0742c2c3a
Spindler, K.D.
6b511593-a50c-40d8-8a8a-bdc6790aaefd
Nieva, C.
a8fc81a0-512a-4bea-863d-c896cf37a2a9
Gwoźdź, T.
ce8ac030-3ae8-4191-bd27-36a2c458ab87
Dutko-Gwoźdź, J.
a3f2ee57-5f8c-46e5-88f0-f9ef16cc9667
Wiedenmann, J.
ad445af2-680f-4927-90b3-589ac9d538f7
Spindler-Barth, M.
1bcdd718-ca1a-4e79-8193-954b6150bd2d
Wieczorek, E.
172d17d7-a902-463c-b6c2-720d26c69537
Dobrucki, J.
615b2dbd-a401-4ae0-9ee4-1114edc47ed6
Duś, D.
44b73782-7c26-4266-a594-e03578dc2313
Henrich, V.
524e2151-9aa4-4004-a474-823d392776db
Ożyhar, A.
f60f3cb9-7823-42c2-8da6-7db0742c2c3a
Spindler, K.D.
6b511593-a50c-40d8-8a8a-bdc6790aaefd

Nieva, C., Gwoźdź, T., Dutko-Gwoźdź, J., Wiedenmann, J., Spindler-Barth, M., Wieczorek, E., Dobrucki, J., Duś, D., Henrich, V., Ożyhar, A. and Spindler, K.D. (2005) Ultraspiracle promotes the nuclear localization of ecdysteroid receptor in mammalian cells. Biological Chemistry, 386 (5), 463-470. (doi:10.1515/BC.2005.055).

Record type: Article

Abstract

The heterodimer consisting of ecdysteroid receptor (EcR) and ultraspiracle (USP), both of which are members of the nuclear receptor superfamily, is considered to be the functional ecdysteroid receptor. Here we analyzed the subcellular distribution of EcR and USP fused to fluorescent proteins. The experiments were carried out in mammalian COS-7, CHO-K1 and HeLa cells to facilitate investigation of the subcellular trafficking of EcR and USP in the absence of endogenous expression of these two receptors. The distribution of USP tagged with a yellow fluorescent protein (YFP-USP) was almost exclusively nuclear in all cell types analyzed. The nuclear localization remained constant for at least 1 day after the first visible signs of expression. In contrast, the intracellular distribution of EcR tagged with a yellow fluorescent protein (YFP-EcR) varied and was dependent on time and cell type, although YFP-EcR alone was also able to partially translocate into the nuclear compartment. Coexpression of YFP-EcR with USP tagged with a cyan fluorescent protein (CFP-USP) resulted in exclusively nuclear localization of both proteins in all cell types analyzed. The USP-induced nuclear localization of YFP-EcR was stable for at least 20 hours. These experiments suggest that USP has a profound effect on the subcellular distribution of EcR.

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More information

Published date: May 2005
Keywords: ecdysone, green fluorescent protein, nuclear receptor, subcellular distribution

Identifiers

Local EPrints ID: 51187
URI: http://eprints.soton.ac.uk/id/eprint/51187
ISSN: 1431-6730
PURE UUID: 16e441bb-e25b-4c24-bdb1-221d5b2c3c62
ORCID for J. Wiedenmann: ORCID iD orcid.org/0000-0003-2128-2943

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Date deposited: 08 May 2008
Last modified: 16 Mar 2024 03:53

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Contributors

Author: C. Nieva
Author: T. Gwoźdź
Author: J. Dutko-Gwoźdź
Author: J. Wiedenmann ORCID iD
Author: M. Spindler-Barth
Author: E. Wieczorek
Author: J. Dobrucki
Author: D. Duś
Author: V. Henrich
Author: A. Ożyhar
Author: K.D. Spindler

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