Induction of zinc finger protein RNF6 auto-ubiquitination for the treatment of myeloma and chronic myeloid leukemia

Zhuang，Haixia1; Ren，Ying2; Mao，Chenyu3; Zhong，Yueya1; Zhang，Zubin2; Cao，Biyin2; Zhang，Yuming4; Huang，Jinqi4; Xu，Guoqiang2; Huang，Zhenqian1; Xu，Yujia1; Mao，Xinliang1

2022-09-01

10.1016/j.jbc.2022.102314

JOURNAL OF BIOLOGICAL CHEMISTRY   影响因子和分区

0021-9258

1083-351X

The zinc finger ubiquitin ligase RNF6 has been proposed as a potential therapeutic target in several cancers, but understanding its molecular mechanism of degradation has been elusive. In the present study, we find that RNF6 is degraded via auto-ubiquitination in a manner dependent on its Really Interesting New Gene (RING) domain. We determine that when the RING domain is deleted (ΔRING) or the core cysteine residues in the zinc finger are mutated (C632S/C635S), the WT protein, but not the ΔRING or mutant RNF6 protein, undergoes polyubiquitination. We also identify USP7 as a deubiquitinase of RNF6 by tandem mass spectrometry. We show that USP7 interacts with RNF6 and abolishes its K48-linked polyubiquitination, thereby preventing its degradation. In contrast, we found a USP7-specific inhibitor promotes RNF6 polyubiquitination, degradation, and cell death. Furthermore, we demonstrate the anti-leukemic drug Nilotinib and anti-myeloma drug Panobinostat (LBH589) induce RNF6 K48-linked polyubiquitination and degradation in both multiple myeloma (MM) and leukemia cells. In agreement with our hypothesis on the mode of RNF6 degradation, we show these drugs promote RNF6 auto-ubiquitination in an in vitro ubiquitination system without other E3 ligases. Consistently, reexpression of RNF6 ablates drug-induced MM and leukemia cell apoptosis. Therefore, our results reveal that RNF6 is a RING E3 ligase that undergoes auto-ubiquitination, which could be abolished by USP7 and induced by anti-cancer drugs. We propose that chemical induction of RNF6 auto-ubiquitination and degradation could be a novel strategy for the treatment of hematological malignancies including MM and leukemia.

auto-ubiquitination leukemia myeloma RNF6 USP7

EI

英语

NI论文

其他

National Natural Science Foundation of China[#81770154];National Natural Science Foundation of China[#81970194];National Natural Science Foundation of China[#82100214];Department of Education of Guangdong Province[2021ZDZX2009];Guangzhou Medical University[JCXKJS2022A05];

20223512639082

Cell death ; Diseases ; Mass spectrometry ; Proteins ; Zinc

Biology:461.9 ; Zinc and Alloys:546.3 ; Chemistry:801 ; Organic Compounds:804.1

BIOLOGY & BIOCHEMISTRY

2-s2.0-85136481849

Scopus

1.Department of Hematology,The First Affiliated Hospital & Guangdong Key Laboratory of Protein Modification and Degradation,School of Basic Medical Sciences,Guangzhou Medical University,Guangzhou,Guangdong,China
2.Department of Pharmacology,College of Pharmaceutical Sciences,Soochow University,Suzhou,Jiangsu,China
3.School of Medicine,Southern University of Science and Technology,Shenzhen,Guangdong,China
4.Department of Hematology,Hematology Research Institute,Affiliated Hospital of Guangdong Medical University,Zhanjiang,Guangdong,China

Zhuang，Haixia,Ren，Ying,Mao，Chenyu,et al. Induction of zinc finger protein RNF6 auto-ubiquitination for the treatment of myeloma and chronic myeloid leukemia[J]. JOURNAL OF BIOLOGICAL CHEMISTRY,2022,298(9).

Zhuang，Haixia.,Ren，Ying.,Mao，Chenyu.,Zhong，Yueya.,Zhang，Zubin.,...&Mao，Xinliang.(2022).Induction of zinc finger protein RNF6 auto-ubiquitination for the treatment of myeloma and chronic myeloid leukemia.JOURNAL OF BIOLOGICAL CHEMISTRY,298(9).

Zhuang，Haixia,et al."Induction of zinc finger protein RNF6 auto-ubiquitination for the treatment of myeloma and chronic myeloid leukemia".JOURNAL OF BIOLOGICAL CHEMISTRY 298.9(2022).