Induction of the human <em>CDC45</em> gene promoter activity by natural compound <em>trans</em>‑resveratrol

Arakawa,Jun; Kondoh,Hikaru; Matsushita,Tokiyoshi; Ogino,Yoko; Asai,Masashi; Tanuma,Sei-Ichi; Uchiumi,Fumiaki

doi:10.3892/mmr.2024.13216

Induction of the human CDC45 gene promoter activity by natural compound trans‑resveratrol

Authors:
- Jun Arakawa
- Hikaru Kondoh
- Tokiyoshi Matsushita
- Yoko Ogino
- Masashi Asai
- Sei-Ichi Tanuma
- Fumiaki Uchiumi
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Affiliations: Department of Gene Regulation, Faculty of Pharmaceutical Sciences, Tokyo University of Science, Noda, Chiba 278‑8510, Japan, Genomic Medicinal Science, Research Institute for Science and Technology, Tokyo University of Science, Noda, Chiba 278‑8510, Japan
Published online on: April 4, 2024 https://doi.org/10.3892/mmr.2024.13216
Article Number: 92
Copyright: © Arakawa et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

GGAA motifs in the human TP53 and HELB gene promoters play a part in responding to trans‑resveratrol (Rsv) in HeLa S3 cells. This sequence is also present in the 5'‑upstream region of the human CDC45 gene, which encodes a component of CMG DNA helicase protein complex. The cells were treated with Rsv (20 µM), then transcripts and the translated protein were analyzed by quantitative RT‑PCR and western blotting, respectively. The results showed that the CDC45 gene and protein expression levels were induced after the treatment. To examine whether they were due to the activation of transcription, a 5'‑upstream 556‑bp of the CDC45 gene was cloned and inserted into a multi‑cloning site of the Luciferase (Luc) expression vector. In the present study, various deletion/point mutation‑introduced Luc expression plasmids were constructed and they were used for the transient transfection assay. The results showed that the GGAA motif, which is included in a putative RELB protein recognizing sequence, plays a part in the promoter activity with response to Rsv in HeLa S3 cells.

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1	Simon AC, Sannino V, Costanzo V and Pellegrini L: Structure of human Cdc45 and implications for CMG helicase function. Nat Commun. 7:116382016. View Article : Google Scholar : PubMed/NCBI
2	Szambowska A, Tessmer I, Prus P, Schlott B, Pospiech H and Grosse F: Cdc45-induced loading of human RPA onto single-stranded DNA. Nucleic Acids Res. 45:3217–3230. 2017.PubMed/NCBI
3	Krastanova I, Sannino V, Amenitsch H, Gileadi O, Pisani FM and Onesti S: Structural and functional insights into the DNA replication factor Cdc45 reveal an evolutionary relationship to the DHH family of phosphoesterases. J Biol Chem. 287:4121–4128. 2012. View Article : Google Scholar : PubMed/NCBI
4	Szambowska A, Tessmer I, Kursula P, Usskilat C, Prus P, Pospiech H and Grosse F: DNA binding properties of human Cdc45 suggest a function as molecular wedge for DNA unwinding. Nucleic Acids Res. 42:2308–2319. 2014. View Article : Google Scholar : PubMed/NCBI
5	Bruck I and Kaplan DL: Cdc45 protein-single-stranded DNA interaction is important for stalling the helicase during replication stress. J Biol Chem. 288:7550–7563. 2013. View Article : Google Scholar : PubMed/NCBI
6	Can G, Kauerhof AC, Macak D and Zegerman P: Helicase subunit Cdc45 targets the checkpoint kinase Rad53 to both replication initiation and elongation complexes after fork stalling. Mol Cell. 73:562–573.e3. 2019. View Article : Google Scholar : PubMed/NCBI
7	Tkáč J, Xu G, Adhikary H, Young JTF, Gallo D, Escribano-Díaz C, Krietsch J, Orthwein A, Munro M, Sol W, et al: HELB is a feedback inhibitor of DNA end resection. Mol Cell. 61:405–418. 2016. View Article : Google Scholar : PubMed/NCBI
8	Guler GD, Liu H, Vaithiyalingam S, Arnett DR, Kremmer E, Chazin WJ and Fanning E: Human DNA helicase B (HDHB) binds to replication protein A and facilitates cellular recovery from replication stress. J Biol Chem. 287:6469–6481. 2012. View Article : Google Scholar : PubMed/NCBI
9	Liu H, Yan P and Fanning E: Human DNA helicase B functions in cellular homologous recombination and stimulates Rad51-mediated 5′-3′ heteroduplex extension in vitro. PLoS One. 10:e01168522015. View Article : Google Scholar : PubMed/NCBI
10	Gerhardt J, Guler GD and Fanning E: Human DNA helicase B interacts with the replication initiation protein Cdc45 and facilitates Cdc45 binding onto chromatin. Exp Cell Res. 334:283–293. 2015. View Article : Google Scholar : PubMed/NCBI
11	Huang J, Zhang J, Bellani MA, Pokharel D, Gichimu J, James RC, Gali H, Ling C, Yan Z, Xu D, et al: Remodeling of interstrand crosslink proximal replisomes is dependent on ATR, FANCM, and FANCD2. Cell Rep. 27:1794–1808.e5. 2019. View Article : Google Scholar : PubMed/NCBI
12	Aparicio T, Guillou E, Coloma J, Montoya G and Méndez J: The human GINS complex associates with Cdc45 and MCM and is essential for DNA replication. Nucleic Acids Res. 37:2087–2095. 2009. View Article : Google Scholar : PubMed/NCBI
13	Köhler C, Koalick D, Fabricius A, Parplys AC, Borgmann K, Pospiech H and Grosse F: Cdc45 is limiting for replication initiation in humans. Cell Cycle. 15:974–985. 2016. View Article : Google Scholar : PubMed/NCBI
14	Chen M, Gutierrez GJ and Ronai ZA: Ubiquitin-recognition protein Ufd1 couples the endoplasmic reticulum (ER) stress response to cell cycle control. Proc Natl Acad Sci USA. 108:9119–9124. 2011. View Article : Google Scholar : PubMed/NCBI
15	Maric M, Mukherjee P, Tatham MH, Hay R and Labib K: Ufd-Npl4 recruit Cdc48 for disassembly of ubiquitylated CMG helicase at the end of chromosome replication. Cell Rep. 18:3033–3042. 2017. View Article : Google Scholar : PubMed/NCBI
16	Uchiumi F, Miyazaki S and Tanuma S: The possible functions of duplicated ets (GGAA) motifs located near transcription start sites of various human genes. Cell Mol Life Sci. 68:2039–2051. 2011. View Article : Google Scholar : PubMed/NCBI
17	FitzGerald PC, Shlyakhtenko A, Mir AA and Vinson C: Clustering of DNA sequences in human promoters. Genome Res. 14:1562–1574. 2004. View Article : Google Scholar : PubMed/NCBI
18	Wei GH, Badis G, Berger MF, Kivioja T, Palin K, Enge M, Bonke M, Jolma A, Varjosalo M, Gehrke AR, et al: Genome-wide analysis of ETS-family DNA-binding in vitro and in vivo. EMBO J. 29:2147–2160. 2010. View Article : Google Scholar : PubMed/NCBI
19	Cheng D, Zhao Y, Wang S, Jia W, Kang J and Zhu J: Human telomerase reverse transcriptase (hTERT) transcription requires Sp1/Sp3 binding to the promoter and a permissive chromatin environment. J Biol Chem. 290:30193–30203. 2015. View Article : Google Scholar : PubMed/NCBI
20	Uchiumi F, Arakawa J, Iwakoshi S, Ishibashi S and Tanuma S: Characterization of the 5′-flanking region of the human DNA helicase B (HELB) gene and its response to trans-resveratrol. Sci Rep. 6:245102016. View Article : Google Scholar : PubMed/NCBI
21	Uchiumi F, Katsuda C, Akui M, Kusaka M, Tanaka M, Asai M and Tanuma SI: Effect of the natural compound trans-resveratrol on human MCM4 gene transcription. Oncol Rep. 44:283–292. 2020. View Article : Google Scholar : PubMed/NCBI
22	Uchiumi F, Shoji K, Sasaki Y, Sasaki M, Sasaki Y, Oyama K, Sugisawa S and Tanuma S: Characterization of the 5′-flanking region of the human TP53 gene and its response to the natural compound, resveratrol. J Biochem. 159:437–447. 2016. View Article : Google Scholar : PubMed/NCBI
23	Tanuma S and Kanai Y: Poly(ADP-ribosyl)ation of chromosomal proteins in the HeLa S3 cell cycle. J Biol Chem. 257:6565–6570. 1982. View Article : Google Scholar : PubMed/NCBI
24	Uchiumi F, Watanabe T, Hasegawa S, Hoshi T, Higami Y and Tanuma S: The effect of resveratrol on the werner syndrome RecQ helicase gene and telomerase activity. Curr Aging Sci. 4:1–7. 2011. View Article : Google Scholar : PubMed/NCBI
25	Uchiumi F, Larsen S and Tanuma S: Application of DEAE-dextran to an efficient gene transfer system. Dextran: Chemical Structure, Application and Potential Side Effects. Figgs GP: Nova Science Publishers; Hauppauge, NY: pp. 143–156. 2014
26	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
27	Takihara Y, Sudo D, Arakawa J, Takahashi M, Sato A, Tanuma S and Uchiumi F: Nicotinamide adenine dinucleotide (NAD⁺) and cell aging. New Research on Cell Aging and Death. Strakoš R and Lorens B: Nova Science Publishers; Hauppauge, NY: pp. 131–158. 2018
28	Igaki H, Nakagawa K, Aoki Y, Ohtomo K, Kukimoto I and Kanda T: Characterization of the bi-directional transcriptional control region between the human UFD1L and CDC45 genes. Biochem Biophys Res Commun. 283:569–576. 2001. View Article : Google Scholar : PubMed/NCBI
29	Uchiumi F, Larsen S and Tanuma S: Biological systems that control transcription of DNA repair and telomere maintenance-associated genes. New Research in Directions in DNA Repair. Chen C: InTech Open; London: pp. 309–325. 2013
30	Martino V, Bianchera A, Reia L, Bussolati O, Fazzina R, Marino F, Montemurro L, Tonelli R, Pession A, Gazzola GC and Sala R: Down regulation of HOXA4, HOXA7, HOXA10, HOXA11 and MEIS1 during monocyte-macrophage differentiation in THP-1 cells. Mol Med Rep. 2:241–244. 2009.PubMed/NCBI
31	Calvo KR, Knoepfler PS, Sykes DB, Pasillas MP and Kamps MP: Meis1a suppresses differentiation by G-CSF and promotes proliferation by SCF: Potential mechanisms of cooperativity with Hox9 in myeloid leukemia. Proc Natl Acad Sci USA. 98:13120–13125. 2001. View Article : Google Scholar : PubMed/NCBI
32	Li T, Morgan MJ, Choksi S, Zhang Y, Kim YS and Liu ZG: MicroRNAs modulate the noncanonical transcription factor NF-kappaB pathway by regulating expression of the kinase IKKalpha during macrophage differentiation. Nat Immunol. 11:799–805. 2010. View Article : Google Scholar : PubMed/NCBI
33	Zhao Z, Hou X, Yin X, Li Y, Duan R, Boyce BF and Yao Z: TNF induction of NF-kB RelB enhances RANKL-induced osteoclastogenesis by promoting inflammatory macrophage differentiation but also limits it through suppression of NFATc1 expression. PLoS One. 10:e01357282015. View Article : Google Scholar : PubMed/NCBI
34	Hamada H, Goto Y, Arakawa J, Murayama E, Ogawa Y, Konno M, Oyama T, Asai M, Sato A, Tanuma S and Uchiumi F: Characterization of the human E2F4 promoter region and its response to 12-O-tetradecanoylphorbol-13-acetate. J Biochem. 166:363–373. 2019. View Article : Google Scholar : PubMed/NCBI
35	Hamada H, Yamamura M, Ohi H, Kobayashi Y, Niwa K, Oyama T, Mano Y, Asai M, Tanuma SI and Uchiumi F: Characterization of the human zinc finger nfx-1-type containing 1 encoding ZNFX1 gene and its response to 12-O-tetradecanoyl-13-acetate in HL-60 cells. Int J Oncol. 55:869–904. 2019.
36	Taneja P, Gu J, Peng R, Carrick R, Uchiumi F, Ott RD, Gustafson E, Podust VN and Fanning E: A dominant-negative mutant of human DNA helicase B blocks the onset of chromosomal DNA replication. J Biol Chem. 277:40853–40861. 2002. View Article : Google Scholar : PubMed/NCBI
37	Hazeslip L, Zafar MK, Chauhan MZ and Byrd AK: Genome maintenance by DNA helicase B. Genes (Basel). 11:5782020. View Article : Google Scholar : PubMed/NCBI
38	Miller TCR, Locke J, Greiwe JF, Diffley JFX and Costa A: Mechanism of head-to-head MCM double-hexamer formation revealed by cryo-EM. Nature. 575:704–710. 2019. View Article : Google Scholar : PubMed/NCBI
39	Douglas ME, Ali FA, Costa A and Diffley JFX: The mechanism of eukaryotic CMG helicase activation. Nature. 555:265–268. 2018. View Article : Google Scholar : PubMed/NCBI
40	Araki H: Molecular mechanisms of DNA replication. DNA Replication, Recombination, and Repair. Hanaoka F and Sugasawa K: Springer; Japan, Tokyo: pp. 3–22. 2016, View Article : Google Scholar
41	Uchiumi F: Transcriptional regulatory systems of the human helicases. Advances in Medicine and Biology. Berhardt LV: Nova Science Publishers; Hauppauge, NY: pp. 115–153. 2020
42	Xiang S, Reed DR and Alexandrow MG: The CMG helicase and cancer: A tumor ‘engine’ and weakness with missing mutations. Oncogene. 42:473–490. 2023. View Article : Google Scholar : PubMed/NCBI
43	Lu Y, Chen X, Liu F, Yu H, Zhang Y, Du K, Nan Y and Huang Q: Systematic pan-cancer analysis identifies CDC45 as having an oncogenic role in human cancers. Oncol Rep. 48:1852022. View Article : Google Scholar : PubMed/NCBI
44	Stefani M, Markus MA, Lin RC, Pinese M, Dawes IW and Morris BJ: The effect of resveratrol on a cell model of human aging. Ann N Y Acad Sci. 1114:407–418. 2007. View Article : Google Scholar : PubMed/NCBI
45	Kaeberlein M: Resveratrol and rapamycin: Are they anti-aging drugs? Bioessays. 32:96–99. 2010. View Article : Google Scholar : PubMed/NCBI
46	Uchiumi F, Arakawa J, Takihara Y, Akui M, Ishibashi S and Tanuma S: The effect of trans-resveratrol on the expression of the human DNA-repair associated genes. Int Mol Med. 3:783–792. 2016.