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Acetylation-Mimic Mutation of TRIM28-Lys304 to Gln Attenuates the Interaction with KRAB-Zinc-Finger Proteins and Affects Gene Expression in Leukemic K562 Cells

TRIM28/KAP1/TIF1β is a crucial epigenetic modifier. Genetic ablation of trim28 is embryonic lethal, although RNAi-mediated knockdown in somatic cells yields viable cells. Reduction in TRIM28 abundance at the cellular or organismal level results in polyphenism. Posttranslational modifications such as...

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Autores principales: Chang, Yao-Jen, Lin, Steven, Kang, Zhi-Fu, Shen, Bin-Jon, Tsai, Wen-Hai, Chen, Wen-Ching, Lu, Hsin-Pin, Su, Yu-Lun, Chou, Shu-Jen, Lin, Shu-Yu, Lin, Sheng-Wei, Huang, Yin-Jung, Wang, Hsin-Hui, Chang, Ching-Jin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10298087/
https://www.ncbi.nlm.nih.gov/pubmed/37372979
http://dx.doi.org/10.3390/ijms24129830
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author Chang, Yao-Jen
Lin, Steven
Kang, Zhi-Fu
Shen, Bin-Jon
Tsai, Wen-Hai
Chen, Wen-Ching
Lu, Hsin-Pin
Su, Yu-Lun
Chou, Shu-Jen
Lin, Shu-Yu
Lin, Sheng-Wei
Huang, Yin-Jung
Wang, Hsin-Hui
Chang, Ching-Jin
author_facet Chang, Yao-Jen
Lin, Steven
Kang, Zhi-Fu
Shen, Bin-Jon
Tsai, Wen-Hai
Chen, Wen-Ching
Lu, Hsin-Pin
Su, Yu-Lun
Chou, Shu-Jen
Lin, Shu-Yu
Lin, Sheng-Wei
Huang, Yin-Jung
Wang, Hsin-Hui
Chang, Ching-Jin
author_sort Chang, Yao-Jen
collection PubMed
description TRIM28/KAP1/TIF1β is a crucial epigenetic modifier. Genetic ablation of trim28 is embryonic lethal, although RNAi-mediated knockdown in somatic cells yields viable cells. Reduction in TRIM28 abundance at the cellular or organismal level results in polyphenism. Posttranslational modifications such as phosphorylation and sumoylation have been shown to regulate TRIM28 activity. Moreover, several lysine residues of TRIM28 are subject to acetylation, but how acetylation of TRIM28 affects its functions remains poorly understood. Here, we report that, compared with wild-type TRIM28, the acetylation-mimic mutant TRIM28-K304Q has an altered interaction with Krüppel-associated box zinc-finger proteins (KRAB-ZNFs). The TRIM28-K304Q knock-in cells were created in K562 erythroleukemia cells by CRISPR-Cas9 (Clustered regularly interspaced short palindromic repeats/CRISPR-associated protein nuclease 9) gene editing method. Transcriptome analysis revealed that TRIM28-K304Q and TRIM28 knockout K562 cells had similar global gene expression profiles, yet the profiles differed considerably from wild-type K562 cells. The expression levels of embryonic-related globin gene and a platelet cell marker integrin-beta 3 were increased in TRIM28-K304Q mutant cells, indicating the induction of differentiation. In addition to the differentiation-related genes, many zinc-finger-proteins genes and imprinting genes were activated in TRIM28-K304Q cells; they were inhibited by wild-type TRIM28 via binding with KRAB-ZNFs. These results suggest that acetylation/deacetylation of K304 in TRIM28 constitutes a switch for regulating its interaction with KRAB-ZNFs and alters the gene regulation as demonstrated by the acetylation mimic TRIM28-K304Q.
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spelling pubmed-102980872023-06-28 Acetylation-Mimic Mutation of TRIM28-Lys304 to Gln Attenuates the Interaction with KRAB-Zinc-Finger Proteins and Affects Gene Expression in Leukemic K562 Cells Chang, Yao-Jen Lin, Steven Kang, Zhi-Fu Shen, Bin-Jon Tsai, Wen-Hai Chen, Wen-Ching Lu, Hsin-Pin Su, Yu-Lun Chou, Shu-Jen Lin, Shu-Yu Lin, Sheng-Wei Huang, Yin-Jung Wang, Hsin-Hui Chang, Ching-Jin Int J Mol Sci Article TRIM28/KAP1/TIF1β is a crucial epigenetic modifier. Genetic ablation of trim28 is embryonic lethal, although RNAi-mediated knockdown in somatic cells yields viable cells. Reduction in TRIM28 abundance at the cellular or organismal level results in polyphenism. Posttranslational modifications such as phosphorylation and sumoylation have been shown to regulate TRIM28 activity. Moreover, several lysine residues of TRIM28 are subject to acetylation, but how acetylation of TRIM28 affects its functions remains poorly understood. Here, we report that, compared with wild-type TRIM28, the acetylation-mimic mutant TRIM28-K304Q has an altered interaction with Krüppel-associated box zinc-finger proteins (KRAB-ZNFs). The TRIM28-K304Q knock-in cells were created in K562 erythroleukemia cells by CRISPR-Cas9 (Clustered regularly interspaced short palindromic repeats/CRISPR-associated protein nuclease 9) gene editing method. Transcriptome analysis revealed that TRIM28-K304Q and TRIM28 knockout K562 cells had similar global gene expression profiles, yet the profiles differed considerably from wild-type K562 cells. The expression levels of embryonic-related globin gene and a platelet cell marker integrin-beta 3 were increased in TRIM28-K304Q mutant cells, indicating the induction of differentiation. In addition to the differentiation-related genes, many zinc-finger-proteins genes and imprinting genes were activated in TRIM28-K304Q cells; they were inhibited by wild-type TRIM28 via binding with KRAB-ZNFs. These results suggest that acetylation/deacetylation of K304 in TRIM28 constitutes a switch for regulating its interaction with KRAB-ZNFs and alters the gene regulation as demonstrated by the acetylation mimic TRIM28-K304Q. MDPI 2023-06-06 /pmc/articles/PMC10298087/ /pubmed/37372979 http://dx.doi.org/10.3390/ijms24129830 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Chang, Yao-Jen
Lin, Steven
Kang, Zhi-Fu
Shen, Bin-Jon
Tsai, Wen-Hai
Chen, Wen-Ching
Lu, Hsin-Pin
Su, Yu-Lun
Chou, Shu-Jen
Lin, Shu-Yu
Lin, Sheng-Wei
Huang, Yin-Jung
Wang, Hsin-Hui
Chang, Ching-Jin
Acetylation-Mimic Mutation of TRIM28-Lys304 to Gln Attenuates the Interaction with KRAB-Zinc-Finger Proteins and Affects Gene Expression in Leukemic K562 Cells
title Acetylation-Mimic Mutation of TRIM28-Lys304 to Gln Attenuates the Interaction with KRAB-Zinc-Finger Proteins and Affects Gene Expression in Leukemic K562 Cells
title_full Acetylation-Mimic Mutation of TRIM28-Lys304 to Gln Attenuates the Interaction with KRAB-Zinc-Finger Proteins and Affects Gene Expression in Leukemic K562 Cells
title_fullStr Acetylation-Mimic Mutation of TRIM28-Lys304 to Gln Attenuates the Interaction with KRAB-Zinc-Finger Proteins and Affects Gene Expression in Leukemic K562 Cells
title_full_unstemmed Acetylation-Mimic Mutation of TRIM28-Lys304 to Gln Attenuates the Interaction with KRAB-Zinc-Finger Proteins and Affects Gene Expression in Leukemic K562 Cells
title_short Acetylation-Mimic Mutation of TRIM28-Lys304 to Gln Attenuates the Interaction with KRAB-Zinc-Finger Proteins and Affects Gene Expression in Leukemic K562 Cells
title_sort acetylation-mimic mutation of trim28-lys304 to gln attenuates the interaction with krab-zinc-finger proteins and affects gene expression in leukemic k562 cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10298087/
https://www.ncbi.nlm.nih.gov/pubmed/37372979
http://dx.doi.org/10.3390/ijms24129830
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