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Functional dissection of N-terminal nuclear trafficking signals of SETDB1

SETDB1 is a histone H3-lysine 9-specific methyltransferase that fulfills epigenetic functions inside the nucleus; however, when overexpressed, SETDB1 majorily localizes in the cytoplasm. SETDB1 has a single nuclear-localization-signal (NLS) motif and two successive nuclear-export-signal (NES1 and NE...

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Autores principales: Eom, Jaemin, Jeon, Kyuheum, Park, Jung Sun, Kang, Yong-Kook
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9807615/
https://www.ncbi.nlm.nih.gov/pubmed/36605719
http://dx.doi.org/10.3389/fcell.2022.1069765
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author Eom, Jaemin
Jeon, Kyuheum
Park, Jung Sun
Kang, Yong-Kook
author_facet Eom, Jaemin
Jeon, Kyuheum
Park, Jung Sun
Kang, Yong-Kook
author_sort Eom, Jaemin
collection PubMed
description SETDB1 is a histone H3-lysine 9-specific methyltransferase that fulfills epigenetic functions inside the nucleus; however, when overexpressed, SETDB1 majorily localizes in the cytoplasm. SETDB1 has a single nuclear-localization-signal (NLS) motif and two successive nuclear-export-signal (NES1 and NES2) motifs in the N-terminus, suggesting that SETDB1 localization is the consequence of a balance between the two antithetic motifs. Here, we performed a series of motif deletions to characterize their effects on the cellular movement of SETDB1. Given the cytoplasmic localization of GFP-SETDB1 in the whole form, without the NES motifs, GFP-SETDB1 was not nuclear, and 3xNLS addition plus NES removal held the majority of GFP-SETDB1 within the nucleus. The results indicated that the cytoplasmic localization of GFP-SETDB1 is the combined result of weak NLS and robust NESs. In ATF7IP-overexpressing cells, GFP-SETDB1 entered the nucleus only in the presence of the NES1 motif; neither the NES2 nor NLS motif was necessary. Since subcellular fractionation results showed that ATF7IP was nuclear-only, an intermediary protein may interact specifically with the NES1 motif after stimulation by ATF7IP. When GFP-SETDB1 had either NES1 or NES2, it was precipitated (in immunoprecipitation) and colocalized (in immunofluorescence) with ATF7IP, indicating that GFP-SETDB1 interacts with ATF7IP through the NES motifs in the nucleus. The regulated nuclear entry of SETDB1 is assumed to set a tight restriction on its abundance within the nucleus, thereby ensuring balanced nuclear SETDB1 levels.
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spelling pubmed-98076152023-01-04 Functional dissection of N-terminal nuclear trafficking signals of SETDB1 Eom, Jaemin Jeon, Kyuheum Park, Jung Sun Kang, Yong-Kook Front Cell Dev Biol Cell and Developmental Biology SETDB1 is a histone H3-lysine 9-specific methyltransferase that fulfills epigenetic functions inside the nucleus; however, when overexpressed, SETDB1 majorily localizes in the cytoplasm. SETDB1 has a single nuclear-localization-signal (NLS) motif and two successive nuclear-export-signal (NES1 and NES2) motifs in the N-terminus, suggesting that SETDB1 localization is the consequence of a balance between the two antithetic motifs. Here, we performed a series of motif deletions to characterize their effects on the cellular movement of SETDB1. Given the cytoplasmic localization of GFP-SETDB1 in the whole form, without the NES motifs, GFP-SETDB1 was not nuclear, and 3xNLS addition plus NES removal held the majority of GFP-SETDB1 within the nucleus. The results indicated that the cytoplasmic localization of GFP-SETDB1 is the combined result of weak NLS and robust NESs. In ATF7IP-overexpressing cells, GFP-SETDB1 entered the nucleus only in the presence of the NES1 motif; neither the NES2 nor NLS motif was necessary. Since subcellular fractionation results showed that ATF7IP was nuclear-only, an intermediary protein may interact specifically with the NES1 motif after stimulation by ATF7IP. When GFP-SETDB1 had either NES1 or NES2, it was precipitated (in immunoprecipitation) and colocalized (in immunofluorescence) with ATF7IP, indicating that GFP-SETDB1 interacts with ATF7IP through the NES motifs in the nucleus. The regulated nuclear entry of SETDB1 is assumed to set a tight restriction on its abundance within the nucleus, thereby ensuring balanced nuclear SETDB1 levels. Frontiers Media S.A. 2022-12-20 /pmc/articles/PMC9807615/ /pubmed/36605719 http://dx.doi.org/10.3389/fcell.2022.1069765 Text en Copyright © 2022 Eom, Jeon, Park and Kang. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Cell and Developmental Biology
Eom, Jaemin
Jeon, Kyuheum
Park, Jung Sun
Kang, Yong-Kook
Functional dissection of N-terminal nuclear trafficking signals of SETDB1
title Functional dissection of N-terminal nuclear trafficking signals of SETDB1
title_full Functional dissection of N-terminal nuclear trafficking signals of SETDB1
title_fullStr Functional dissection of N-terminal nuclear trafficking signals of SETDB1
title_full_unstemmed Functional dissection of N-terminal nuclear trafficking signals of SETDB1
title_short Functional dissection of N-terminal nuclear trafficking signals of SETDB1
title_sort functional dissection of n-terminal nuclear trafficking signals of setdb1
topic Cell and Developmental Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9807615/
https://www.ncbi.nlm.nih.gov/pubmed/36605719
http://dx.doi.org/10.3389/fcell.2022.1069765
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