Plk1 interacts with RNF2 and promotes its ubiquitin‑dependent degradation

An,Ran; Cheng,Li; Chen,Lijian; Du,Jian

doi:10.3892/or.2018.6326

Plk1 interacts with RNF2 and promotes its ubiquitin‑dependent degradation

Authors:
- Ran An
- Li Cheng
- Lijian Chen
- Jian Du
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Affiliations: Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Anhui Medical University, Hefei, Anhui 230032, P.R. China, Department of Orthopaedics, The Second Affiliated Hospital of Anhui Medical University, Hefei, Anhui 230032, P.R. China, Department of Anesthesiology, The First Affiliated Hospital, Anhui Medical University, Hefei, Anhui 230032, P.R. China, Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Anhui Medical University, Hefei, Anhui 230032, P.R. China
Published online on: March 20, 2018 https://doi.org/10.3892/or.2018.6326
Pages: 2358-2364

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Abstract

Ring finger protein 2 (RNF2), also known as RING2 or RING1B, displays oncogenic functions in different types of cancers, yet, the function of RNF2 during mitosis has not been evaluated. A yeast two‑hybrid screen was undertaken using a human HeLa cDNA library to explore and identify proteins that interact with RNF2. Several positive clones, including Polo‑like kinase 1 (Plk1), a critical regulator of mitosis, were identified. The interaction between RNF2 and Plk1 was confirmed using a β‑galactosidase and growth test in selective media, in vitro glutathione S‑transferase pull‑down, and in vivo immunoprecipitation assays. Moreover, we confirmed that RNF2 co‑localized with Plk1 at mitotic chromosomes in the prometaphase and metaphase using an immunoﬂuorescence assay. In addition, our results revealed that Plk1 kinase activity was required for ubiquitin‑dependent degradation of RNF2. These findings provide a new clue for understanding the function of RNF2 during mitotic regulation and tumorigenesis.

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1	Zhang Z, Chen C, Cui P, Liao Y, Yao L, Zhang Y, Rui R and Ju S: Plk1 inhibition leads to a failure of mitotic division during the first mitotic division in pig embryos. J Assist Reprod Genet. 34:399–407. 2017. View Article : Google Scholar : PubMed/NCBI
2	Liu J and Zhang C: The equilibrium of ubiquitination and deubiquitination at PLK1 regulates sister chromatid separation. Cell Mol Life Sci. 74:2127–2134. 2017. View Article : Google Scholar : PubMed/NCBI
3	Liu D, Davydenko O and Lampson MA: Polo-like kinase-1 regulates kinetochore-microtubule dynamics and spindle checkpoint silencing. J Cell Biol. 198:491–499. 2012. View Article : Google Scholar : PubMed/NCBI
4	Archambault V, Lépine G and Kachaner D: Understanding the polo kinase machine. Oncogene. 34:4799–4807. 2015. View Article : Google Scholar : PubMed/NCBI
5	Chen F, Zhuo X, Qin T, Guo X, Zhang C and Lai L: Designed inhibitor for nuclear localization signal of polo-like kinase 1 induces mitotic arrest. Chem Biol Drug Des. 89:732–740. 2017. View Article : Google Scholar : PubMed/NCBI
6	Jang YJ, Ma S, Terada Y and Erikson RL: Phosphorylation of threonine 210 and the role of serine 137 in the regulation of mammalian polo-like kinase. J Biol Chem. 277:44115–44120. 2002. View Article : Google Scholar : PubMed/NCBI
7	Gheghiani L, Loew D, Lombard B, Mansfeld J and Gavet O: PLK1 activation in late G2 sets up commitment to mitosis. Cell Rep. 19:2060–2073. 2017. View Article : Google Scholar : PubMed/NCBI
8	Kishi K, van Vugt MA, Okamoto K, Hayashi Y and Yaffe MB: Functional dynamics of Polo-like kinase 1 at the centrosome. Mol Cell Biol. 29:3134–3150. 2009. View Article : Google Scholar : PubMed/NCBI
9	Sumara I, Giménez-Abián JF, Gerlich D, Hirota T, Kraft C, de la Torre C, Ellenberg J and Peters JM: Roles of polo-like kinase 1 in the assembly of functional mitotic spindles. Curr Biol. 14:1712–1722. 2004. View Article : Google Scholar : PubMed/NCBI
10	Carmena M, Pinson X, Platani M, Salloum Z, Xu Z, Clark A, Macisaac F, Ogawa H, Eggert U, Glover DM, et al: The chromosomal passenger complex activates Polo kinase at centromeres. PLoS Biol. 10:e10012502012. View Article : Google Scholar : PubMed/NCBI
11	Bentley ML, Corn JE, Dong KC, Phung Q, Cheung TK and Cochran AG: Recognition of UbcH5c and the nucleosome by the Bmi1/Ring1b ubiquitin ligase complex. EMBO J. 30:3285–3297. 2011. View Article : Google Scholar : PubMed/NCBI
12	Qian T, Lee JY, Park JH, Kim HJ and Kong G: Id1 enhances RING1b E3 ubiquitin ligase activity through the Mel-18/Bmi-1 polycomb group complex. Oncogene. 29:5818–5827. 2010. View Article : Google Scholar : PubMed/NCBI
13	Ben-Saadon R, Zaaroor D, Ziv T and Ciechanover A: The polycomb protein Ring1B generates self atypical mixed ubiquitin chains required for its in vitro histone H2A ligase activity. Mol Cell. 24:701–711. 2006. View Article : Google Scholar : PubMed/NCBI
14	Wen W, Peng C, Kim MO, Jeong Ho C, Zhu F, Yao K, Zykova T, Ma W, Carper A, Langfald A, et al: Knockdown of RNF2 induces apoptosis by regulating MDM2 and p53 stability. Oncogene. 33:421–428. 2014. View Article : Google Scholar : PubMed/NCBI
15	Su WJ, Fang JS, Cheng F, Liu C, Zhou F and Zhang J: RNF2/Ring1b negatively regulates p53 expression in selective cancer cell types to promote tumor development. Proc Natl Acad Sci USA. 110:1720–1725. 2013. View Article : Google Scholar : PubMed/NCBI
16	Wei M, Jiao D, Han D, Wu J, Wei F, Zheng G, Guo Z, Xi W, Yang F, Xie P, et al: Knockdown of RNF2 induces cell cycle arrest and apoptosis in prostate cancer cells through the upregulation of TXNIP. Oncotarget. 8:5323–5338. 2017.PubMed/NCBI
17	Qu C and Qu Y: Down-regulation of salt-inducible kinase 1 (SIK1) is mediated by RNF2 in hepatocarcinogenesis. Oncotarget. 8:3144–3155. 2017. View Article : Google Scholar : PubMed/NCBI
18	Du J, An R, Chen L, Shen Y, Chen Y, Cheng L, Jiang Z, Zhang A, Yu L, Chu D, et al: Toxoplasma gondii virulence factor ROP18 inhibits the host NF-κB pathway by promoting p65 degradation. J Biol Chem. 289:12578–12592. 2014. View Article : Google Scholar : PubMed/NCBI
19	Zhen CY, Duc HN, Kokotovic M, Phiel CJ and Ren X: Cbx2 stably associates with mitotic chromosomes via a PRC2- or PRC1-independent mechanism and is needed for recruiting PRC1 complex to mitotic chromosomes. Mol Biol Cell. 25:3726–3739. 2014. View Article : Google Scholar : PubMed/NCBI
20	Zhang Y, Li X, Chen Z and Bepler G: Ubiquitination and degradation of ribonucleotide reductase M1 by the polycomb group proteins RNF2 and Bmi1 and cellular response to gemcitabine. PLoS One. 9:e911862014. View Article : Google Scholar : PubMed/NCBI
21	Steffen PA, Fonseca JP, Gänger C, Dworschak E, Kockmann T, Beisel C and Ringrose L: Quantitative in vivo analysis of chromatin binding of Polycomb and Trithorax group proteins reveals retention of ASH1 on mitotic chromatin. Nucleic Acids Res. 41:5235–5250. 2013. View Article : Google Scholar : PubMed/NCBI
22	Fonseca JP, Steffen PA, Müller S, Lu J, Sawicka A, Seiser C and Ringrose L: In vivo Polycomb kinetics and mitotic chromatin binding distinguish stem cells from differentiated cells. Genes Dev. 26:857–871. 2012. View Article : Google Scholar : PubMed/NCBI
23	Fukushima H, Ogura K, Wan L, Lu Y, Li V, Gao D, Liu P, Lau AW, Wu T, Kirschner MW, et al: SCF-mediated Cdh1 degradation defines a negative feedback system that coordinates cell-cycle progression. Cell Rep. 4:803–816. 2013. View Article : Google Scholar : PubMed/NCBI
24	Lee YJ, Kim EH, Lee JS, Jeoung D, Bae S, Kwon SH and Lee YS: HSF1 as a mitotic regulator: Phosphorylation of HSF1 by Plk1 is essential for mitotic progression. Cancer Res. 68:7550–7560. 2008. View Article : Google Scholar : PubMed/NCBI
25	Seki A, Coppinger JA, Du H, Jang CY, Yates JR III and Fang G: Plk1- and beta-TrCP-dependent degradation of Bora controls mitotic progression. J Cell Biol. 181:65–78. 2008. View Article : Google Scholar : PubMed/NCBI
26	Li XQ, He WP, Hou WH, Chen JW, Fan RR, Yuan LJ, Yang GP, Cai MY, Chen L, Li J, et al: Overexpression of RNF2 is positively associated with ovarian carcinoma aggressiveness and indicative of poor patient survival. Oncotarget. 6:31181–31190. 2016.
27	Xia P, Wang S, Huang G, Du Y, Zhu P, Li M and Fan Z: RNF2 is recruited by WASH to ubiquitinate AMBRA1 leading to downregulation of autophagy. Cell Res. 24:943–958. 2014. View Article : Google Scholar : PubMed/NCBI
28	Lera RF, Potts GK, Suzuki A, Johnson JM, Salmon ED, Coon JJ and Burkard ME: Decoding Polo-like kinase 1 signaling along the kinetochore-centromere axis. Nat Chem Biol. 12:411–418. 2016. View Article : Google Scholar : PubMed/NCBI
29	Lu LY and Yu X: The balance of Polo-like kinase 1 in tumorigenesis. Cell Div. 4:42009. View Article : Google Scholar : PubMed/NCBI
30	Wachowicz P, Fernández-Miranda G, Marugán C, Escobar B and de Cárcer G: Genetic depletion of Polo-like kinase 1 leads to embryonic lethality due to mitotic aberrancies. Bioessays. 38 Suppl 1:S96–S106. 2016. View Article : Google Scholar : PubMed/NCBI
31	Hasegawa H, Hyodo T, Asano E, Ito S, Maeda M, Kuribayashi H, Natsume A, Wakabayashi T, Hamaguchi M and Senga T: The role of PLK1-phosphorylated SVIL in myosin II activation and cytokinetic furrowing. J Cell Sci. 126:3627–3637. 2013. View Article : Google Scholar : PubMed/NCBI
32	van Vugt MA, Gardino AK, Linding R, Ostheimer GJ, Reinhardt HC, Ong SE, Tan CS, Miao H, Keezer SM, Li J, et al: A mitotic phosphorylation feedback network connects Cdk1, Plk1, 53BP1, and Chk2 to inactivate the G₂/M DNA damage checkpoint. PLoS Biol. 8:e10002872010. View Article : Google Scholar : PubMed/NCBI
33	Lénárt P, Petronczki M, Steegmaier M, Di Fiore B, Lipp JJ, Hoffmann M, Rettig WJ, Kraut N and Peters JM: The small-molecule inhibitor BI 2536 reveals novel insights into mitotic roles of polo-like kinase 1. Curr Biol. 17:304–315. 2007. View Article : Google Scholar : PubMed/NCBI
34	Peng B, Shi R, Jiang W, Ding YH, Dong MQ, Zhu WG and Xu X: Phosphorylation of LSD1 by PLK1 promotes its chromatin release during mitosis. Cell Biosci. 7:152017. View Article : Google Scholar : PubMed/NCBI
35	Park JE, Soung NK, Johmura Y, Kang YH, Liao C, Lee KH, Park CH, Nicklaus MC and Lee KS: Polo-box domain: A versatile mediator of polo-like kinase function. Cell Mol Life Sci. 67:1957–1970. 2010. View Article : Google Scholar : PubMed/NCBI
36	Taniguchi E, Toyoshima-Morimoto F and Nishida E: Nuclear translocation of plk1 mediated by its bipartite nuclear localization signal. J Biol Chem. 277:48884–48888. 2002. View Article : Google Scholar : PubMed/NCBI
37	Tao YF, Li ZH, Du WW, Xu LX, Ren JL, Li XL, Fang F, Xie Y, Li M, Qian GH, et al: Inhibiting PLK1 induces autophagy of acute myeloid leukemia cells via mammalian target of rapamycin pathway dephosphorylation. Oncol Rep. 37:1419–1429. 2017. View Article : Google Scholar : PubMed/NCBI