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 <i>trim28</i> 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 mo...
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MDPI AG
2023-06-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/12/9830 |
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| author | Yao-Jen Chang Steven Lin Zhi-Fu Kang Bin-Jon Shen Wen-Hai Tsai Wen-Ching Chen Hsin-Pin Lu Yu-Lun Su Shu-Jen Chou Shu-Yu Lin Sheng-Wei Lin Yin-Jung Huang Hsin-Hui Wang Ching-Jin Chang |
| author_facet | Yao-Jen Chang Steven Lin Zhi-Fu Kang Bin-Jon Shen Wen-Hai Tsai Wen-Ching Chen Hsin-Pin Lu Yu-Lun Su Shu-Jen Chou Shu-Yu Lin Sheng-Wei Lin Yin-Jung Huang Hsin-Hui Wang Ching-Jin Chang |
| author_sort | Yao-Jen Chang |
| collection | DOAJ |
| description | TRIM28/KAP1/TIF1β is a crucial epigenetic modifier. Genetic ablation of <i>trim28</i> 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 <i>TRIM28</i>-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 <i>TRIM28</i>-K304Q and <i>TRIM28</i> 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 <i>TRIM28</i>-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 <i>TRIM28</i>-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. |
| first_indexed | 2024-03-11T02:22:48Z |
| format | Article |
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| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-11T02:22:48Z |
| publishDate | 2023-06-01 |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-bf9413bf0ba14d279b341c9029bcadf62023-11-18T10:44:45ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-06-012412983010.3390/ijms24129830Acetylation-Mimic Mutation of TRIM28-Lys304 to Gln Attenuates the Interaction with KRAB-Zinc-Finger Proteins and Affects Gene Expression in Leukemic K562 CellsYao-Jen Chang0Steven Lin1Zhi-Fu Kang2Bin-Jon Shen3Wen-Hai Tsai4Wen-Ching Chen5Hsin-Pin Lu6Yu-Lun Su7Shu-Jen Chou8Shu-Yu Lin9Sheng-Wei Lin10Yin-Jung Huang11Hsin-Hui Wang12Ching-Jin Chang13Institute of Biological Chemistry, Academia Sinica, Taipei 11529, TaiwanInstitute of Biological Chemistry, Academia Sinica, Taipei 11529, TaiwanGraduate Institute of Biochemical Sciences, College of Life Science, National Taiwan University, Taipei 10617, TaiwanInstitute of Biological Chemistry, Academia Sinica, Taipei 11529, TaiwanInstitute of Biological Chemistry, Academia Sinica, Taipei 11529, TaiwanGraduate Institute of Biochemical Sciences, College of Life Science, National Taiwan University, Taipei 10617, TaiwanGraduate Institute of Biochemical Sciences, College of Life Science, National Taiwan University, Taipei 10617, TaiwanInstitute of Biological Chemistry, Academia Sinica, Taipei 11529, TaiwanInstitute of Plant and Microbial Biology, Academia Sinica, Taipei 11529, TaiwanInstitute of Biological Chemistry, Academia Sinica, Taipei 11529, TaiwanInstitute of Biological Chemistry, Academia Sinica, Taipei 11529, TaiwanDepartment of Pediatrics, Division of Pediatric Immunology and Nephrology, Taipei Veterans General Hospital, Taipei 11217, TaiwanDepartment of Pediatrics, Division of Pediatric Immunology and Nephrology, Taipei Veterans General Hospital, Taipei 11217, TaiwanInstitute of Biological Chemistry, Academia Sinica, Taipei 11529, TaiwanTRIM28/KAP1/TIF1β is a crucial epigenetic modifier. Genetic ablation of <i>trim28</i> 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 <i>TRIM28</i>-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 <i>TRIM28</i>-K304Q and <i>TRIM28</i> 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 <i>TRIM28</i>-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 <i>TRIM28</i>-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.https://www.mdpi.com/1422-0067/24/12/9830acetylationCRISPR/Cas9TRIM28KRAB-ZNFK562 |
| spellingShingle | Yao-Jen Chang Steven Lin Zhi-Fu Kang Bin-Jon Shen Wen-Hai Tsai Wen-Ching Chen Hsin-Pin Lu Yu-Lun Su Shu-Jen Chou Shu-Yu Lin Sheng-Wei Lin Yin-Jung Huang Hsin-Hui Wang Ching-Jin Chang Acetylation-Mimic Mutation of TRIM28-Lys304 to Gln Attenuates the Interaction with KRAB-Zinc-Finger Proteins and Affects Gene Expression in Leukemic K562 Cells International Journal of Molecular Sciences acetylation CRISPR/Cas9 TRIM28 KRAB-ZNF K562 |
| 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 | acetylation CRISPR/Cas9 TRIM28 KRAB-ZNF K562 |
| url | https://www.mdpi.com/1422-0067/24/12/9830 |
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