Skip to main content
SpringerLink
Log in

Down-Regulated CUEDC2 Increases GDNF Expression by Stabilizing CREB Through Reducing Its Ubiquitination in Glioma

  • Original Paper
  • Published:
Neurochemical Research Aims and scope Submit manuscript

Abstract

Abnormally high expression of glial cell line-derived neurotrophic factor (GDNF) derived from glioma cells has essential impacts on gliomagenesis and development, but the molecular basis underlying increased GDNF expression in glioma cells remain unclear. This work aimed to study the molecular mechanisms that may explain the accumulation of GDNF in glioma. Firstly, we observed that cAMP response element-binding protein (CREB), known as an important transcription factor for binding of GDNF promoter region, was highly expressed with an apparent accumulation into the nucleus of glioma cells, which may contribute to the transcription of GDNF. Secondly, CUE domain-containing protein 2 (CUEDC2), a ubiquitin-regulated protein, could increase the amount of binding between the E3 ligase tripartite motif-containing 21 (TRIM21) and CREB and affect the CREB level. Like our previous study, it showed that there was a significantly down-regulation of CUEDC2 in glioma. Finally, our data suggest that GDNF expression is indirectly regulated by transcription factor ubiquitination. Indeed, down-regulation of CUEDC2, decreased the ubiquitination and degradation of CREB, which was associated to high levels of GDNF. Furthermore, abundant CREB involved in the binding to the GDNF promoter region contributes to GDNF high expression in glioma cells. Collectively, it was verified the GDNF expression was affected by CREB ubiquitination regulated by CUEDC2 level.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

Abbreviations

GDNF:

Glial cell line-derived neurotrophic factor

CUEDC2:

CUE domain-containing protein 2

TRIM21:

Tripartite motif-containing 21

CREB:

CAMP response element-binding protein

ChIP-PCR:

Chromatin immunoprecipitation-polymerase chain reaction

HA:

Human astrocytes

References

  1. Lin LF, Doherty DH, Lile JD, Bektesh S, Collins F (1993) GDNF: a glial cell line-derived neurotrophic factor for midbrain dopaminergic neurons. Science 260(5111):1130–1132

    Article  CAS  Google Scholar 

  2. Ku MC, Wolf SA, Respondek D, Matyash V, Pohlmann A, Waiczies S, Waiczies H, Niendorf T, Synowitz M, Glass R, Kettenmann H (2013) GDNF mediates glioblastoma-induced microglia attraction but not astrogliosis. Acta Neuropathol 125(4):609–620. https://doi.org/10.1007/s00401-013-1079-8

    Article  CAS  PubMed  Google Scholar 

  3. Lu DY, Leung YM, Cheung CW, Chen YR, Wong KL (2010) Glial cell line-derived neurotrophic factor induces cell migration and matrix metalloproteinase-13 expression in glioma cells. Biochem Pharmacol 80(8):1201–1209. https://doi.org/10.1016/j.bcp.2010.06.046

    Article  CAS  PubMed  Google Scholar 

  4. Ng WH, Wan GQ, Peng ZN, Too HP (2009) Glial cell-line derived neurotrophic factor (GDNF) family of ligands confer chemoresistance in a ligand-specific fashion in malignant gliomas. J Clin Neurosci 16(3):427–436. https://doi.org/10.1016/j.jocn.2008.06.002

    Article  CAS  PubMed  Google Scholar 

  5. Wiesenhofer B, Stockhammer G, Kostron H, Maier H, Hinterhuber H, Humpel C (2000) Glial cell line-derived neurotrophic factor (GDNF) and its receptor (GFR-alpha 1) are strongly expressed in human gliomas. Acta Neuropathol 99(2):131–137

    Article  CAS  Google Scholar 

  6. Yu ZQ, Zhang BL, Ren QX, Wang JC, Yu RT, Qu DW, Liu ZH, Xiong Y, Gao DS (2013) Changes in transcriptional factor binding capacity resulting from promoter region methylation induce aberrantly high GDNF expression in human glioma. Mol Neurobiol 48(3):571–580. https://doi.org/10.1007/s12035-013-8443-5

    Article  CAS  PubMed  Google Scholar 

  7. Zhang L, Wang D, Han X, Tang F, Gao D (2019) Mechanism of methylation and acetylation of high GDNF transcription in glioma cells: a review. Heliyon 5(6):e01951

    Article  Google Scholar 

  8. Zhang B, Gu X, Han X, Gao Q, Liu J, Guo T, Gao D (2020) Crosstalk between DNA methylation and histone acetylation triggers GDNF high transcription in glioblastoma cells. Clin Epigenetics 12:47. https://doi.org/10.1186/s13148-020-00835-3

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Firestein R, Feuerstein N (1998) Association of activating transcription factor 2 (ATF2) with the ubiquitin-conjugating enzyme hUBC9. Implication of the ubiquitin/proteasome pathway in regulation of ATF2 in T cells. J Biol Chem 273(10):5892–5902

    Article  CAS  Google Scholar 

  10. Fuchs SY, Ronai Z (1999) Ubiquitination and degradation of ATF2 are dimerization dependent. Mol Cell Biol 19(5):3289–3298

    Article  CAS  Google Scholar 

  11. Sutter CH, Laughner E, Semenza GL (2000) Hypoxia-inducible factor 1alpha protein expression is controlled by oxygen-regulated ubiquitination that is disrupted by deletions and missense mutations. Proc Natl Acad Sci USA 97(9):4748–4753. https://doi.org/10.1073/pnas.080072497

    Article  CAS  PubMed  Google Scholar 

  12. Yaron A, Hatzubai A, Davis M, Lavon I, Amit S, Manning AM, Andersen JS, Mann M, Mercurio F, Ben-Neriah Y (1998) Identification of the receptor component of the IkappaBalpha-ubiquitin ligase. Nature 396(6711):590–594. https://doi.org/10.1038/25159

    Article  CAS  PubMed  Google Scholar 

  13. Taylor CT, Furuta GT, Synnestvedt K, Colgan SP (2000) Phosphorylation-dependent targeting of cAMP response element binding protein to the ubiquitin/proteasome pathway in hypoxia. Proc Natl Acad Sci USA 97(22):12091–12096. https://doi.org/10.1073/pnas.220211797

    Article  CAS  PubMed  Google Scholar 

  14. Zhang PJ, Zhao J, Li HY, Man JH, He K, Zhou T, Pan X, Li AL, Gong WL, Jin BF, Xia Q, Yu M, Shen BF, Zhang XM (2007) CUE domain containing 2 regulates degradation of progesterone receptor by ubiquitin-proteasome. EMBO J 26(7):1831–1842. https://doi.org/10.1038/sj.emboj.7601602

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Donaldson KM, Yin H, Gekakis N, Supek F, Joazeiro CA (2003) Ubiquitin signals protein trafficking via interaction with a novel ubiquitin binding domain in the membrane fusion regulator, Vps9p. Curr Biol CB 13(3):258–262

    Article  CAS  Google Scholar 

  16. Man J, Zhang X (2011) CUEDC2: an emerging key player in inflammation and tumorigenesis. Protein Cell 2(9):699–703. https://doi.org/10.1007/s13238-011-1089-z

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Jian Z, Liang B, Pan X, Xu G, Guo SS, Li T, Zhou T, Xiao YB (2016) CUEDC2 modulates cardiomyocyte oxidative capacity by regulating GPX1 stability. EMBO Mol Med 8(7):813–829. https://doi.org/10.15252/emmm.201506010

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Li F, Tang C, Jin D, Guan L, Wu Y, Liu X, Wu X, Wu QY, Gao D (2017) CUEDC2 suppresses glioma tumorigenicity by inhibiting the activation of STAT3 and NF-kappaB signaling pathway. Int J Oncol 51(1):115–127. https://doi.org/10.3892/ijo.2017.4009

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Woodbury D, Schaar DG, Ramakrishnan L, Black IBJBR (1998) Novel structure of the human GDNF gene. Brain Res 803(1–2):95–104

    Article  CAS  Google Scholar 

  20. Wan G, Too HP (2010) A specific isoform of glial cell line-derived neurotrophic factor family receptor alpha 1 regulates RhoA expression and glioma cell migration. J Neurochem 115(3):759–770. https://doi.org/10.1111/j.1471-4159.2010.06975.x

    Article  CAS  PubMed  Google Scholar 

  21. Xiong Y, Liu L, Zhu S, Zhang B, Qin Y, Yao R, Zhou H, Gao DS (2017) Precursor N-cadherin mediates glial cell line-derived neurotrophic factor-promoted human malignant glioma. Oncotarget 8(15):24902–24914. https://doi.org/10.18632/oncotarget.15302

    Article  PubMed  PubMed Central  Google Scholar 

  22. Montminy MR, Bilezikjian LM (1987) Binding of a nuclear protein to the cyclic-AMP response element of the somatostatin gene. Nature 328(6126):175–178. https://doi.org/10.1038/328175a0

    Article  CAS  Google Scholar 

  23. Yamamoto KK, Gonzalez GA, Biggs WH 3rd, Montminy MR (1988) Phosphorylation-induced binding and transcriptional efficacy of nuclear factor CREB. Nature 334(6182):494–498. https://doi.org/10.1038/334494a0

    Article  CAS  PubMed  Google Scholar 

  24. Mayr B, Montminy M (2001) Transcriptional regulation by the phosphorylation-dependent factor CREB. Nat Rev Mol Cell Biol 2(8):599–609. https://doi.org/10.1038/35085068

    Article  CAS  PubMed  Google Scholar 

  25. Shaywitz AJ, Greenberg ME (1999) CREB: a stimulus-induced transcription factor activated by a diverse array of extracellular signals. Annu Rev Biochem 68:821–861. https://doi.org/10.1146/annurev.biochem.68.1.821

    Article  CAS  PubMed  Google Scholar 

  26. Shukla A, Bosenberg MW, MacPherson MB, Butnor KJ, Heintz NH, Pass HI, Carbone M, Testa JR, Mossman BT (2009) Activated cAMP response element binding protein is overexpressed in human mesotheliomas and inhibits apoptosis. Am J Pathol 175(5):2197–2206. https://doi.org/10.2353/ajpath.2009.090400

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Golan M, Schreiber G, Avissar S (2011) Antidepressants elevate GDNF expression and release from C(6) glioma cells in a beta-arrestin1-dependent, CREB interactive pathway. Int J Neuropsychopharmacol 14(10):1289–1300. https://doi.org/10.1017/s1461145710001550

    Article  CAS  PubMed  Google Scholar 

  28. Hisaoka K, Maeda N, Tsuchioka M, Takebayashi M (2008) Antidepressants induce acute CREB phosphorylation and CRE-mediated gene expression in glial cells: a possible contribution to GDNF production. Brain Res 1196:53–58. https://doi.org/10.1016/j.brainres.2007.12.019

    Article  CAS  PubMed  Google Scholar 

  29. Daniel P, Filiz G, Brown DV, Hollande F, Gonzales M, D'Abaco G, Papalexis N, Phillips WA, Malaterre J, Ramsay RG, Mantamadiotis T (2014) Selective CREB-dependent cyclin expression mediated by the PI3K and MAPK pathways supports glioma cell proliferation. Oncogenesis 3:e108. https://doi.org/10.1038/oncsis.2014.21

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Perry C, Sklan EH, Soreq H (2004) CREB regulates AChE-R-induced proliferation of human glioblastoma cells. Neoplasia 6(3):279–286. https://doi.org/10.1593/neo.3424

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Tan X, Wang S, Yang B, Zhu L, Yin B, Chao T, Zhao J, Yuan J, Qiang B, Peng X (2012) The CREB-miR-9 negative feedback minicircuitry coordinates the migration and proliferation of glioma cells. PLoS ONE 7(11):e49570. https://doi.org/10.1371/journal.pone.0049570

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Tan X, Wang S, Zhu L, Wu C, Yin B, Zhao J, Yuan J, Qiang B, Peng X (2012) cAMP response element-binding protein promotes gliomagenesis by modulating the expression of oncogenic microRNA-23a. Proc Natl Acad Sci USA 109(39):15805–15810. https://doi.org/10.1073/pnas.1207787109

    Article  Google Scholar 

  33. Lee JH, Liu R, Li J, Zhang C, Wang Y, Cai Q, Qian X, Xia Y, Zheng Y, Piao Y, Chen Q, de Groot JF, Jiang T, Lu Z (2017) Stabilization of phosphofructokinase 1 platelet isoform by AKT promotes tumorigenesis. Nat Commun 8(1):949. https://doi.org/10.1038/s41467-017-00906-9

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Foltz C, Napolitano A, Khan R, Clough B, Hirst EM, Frickel EM (2017) TRIM21 is critical for survival of Toxoplasma gondii infection and localises to GBP-positive parasite vacuoles. Sci Rep 7(1):5209. https://doi.org/10.1038/s41598-017-05487-7

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Zhao Z, Wang Y, Yun D, Huang Q, Meng D, Li Q, Zhang P, Wang C, Chen H, Lu D (2020) TRIM21 overexpression promotes tumor progression by regulating cell proliferation, cell migration and cell senescence in human glioma. Am J Cancer Res 10(1):114–130

    CAS  PubMed  PubMed Central  Google Scholar 

  36. Müller J, Maurer V, Reimers K, Vogt P, Bucan V (2015) TRIM21, a negative modulator of LFG in breast carcinoma MDA-MB-231 cells in vitro. Int J Oncol 47(5):1634–1646

    Article  Google Scholar 

  37. Si W, Zhou J, Zhao Y, Zheng J, Cui L (2020) SET7/9 promotes multiple malignant processes in breast cancer development via RUNX2 activation and is negatively regulated by TRIM21. Cell Death Dis 11(2):151. https://doi.org/10.1038/s41419-020-2350-2

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Zhou W, Zhang Y, Zhong C, Hu J, Hu H, Zhou D, Cao M (2018) Decreased expression of TRIM21 indicates unfavorable outcome and promotes cell growth in breast cancer. Cancer Manag Res 10:3687–3696. https://doi.org/10.2147/CMAR.S175470

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We are grateful for glioma samples provided by the Affiliated Hospital of Xuzhou Medical University. We would like to acknowledge the Department of Anatomy and Neurobiology of Xuzhou Medical University.

Funding

This study was funded by the National Natural Science Research Foundation of China (nos. 81602464, 81772688), Six Talent Peaks in Jiangsu Province Jiangsu (SWYY-088), Qing Lan Project in Jiangsu Province (2017 to B.L. Zhang), and a Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD).

Author information

Author notes

  1. Xin-Feng Liu and Chuan-Xi Tang have contributed equally to this work.

Authors and Affiliations

  1. Department of Neurobiology and Anatomy, Xuzhou Key Laboratory of Neurobiology, Xuzhou Medical University, 209 Tongshan Road, Xuzhou, 221004, Jiangsu, China

    Xin-Feng Liu, Chuan-Xi Tang, Lin Zhang, Shu-Yan Tong, Yue Wang, Ayanlaja Abiola Abdulrahman, Guang-Quan Ji, Yue Gao, Dian-shuai Gao & Bao-Le Zhang

  2. Department of Emergency Surgery, Xuzhou First People’s Hospital, The Affiliated Xuzhou Municipal Hospital of Xuzhou Medical University, Xuzhou, 221000, Jiangsu, China

    Xin-Feng Liu

Authors
  1. Xin-Feng Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Chuan-Xi Tang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Lin Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Shu-Yan Tong
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yue Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Ayanlaja Abiola Abdulrahman
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Guang-Quan Ji
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Yue Gao
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Dian-shuai Gao
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Bao-Le Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Z-BL and D-SG made substantial contributions to conception and design, funding, and project supervision. X-FL and C-XT conceived the study, performed most experiments, analyzed data, and wrote the manuscript. LZ and S-YT performed and checked the experiments, integrated data. AA Abdulrahman embellished the paper. YW, G-QJ and YG were responsible for statistical analysis and organizing pictures and tables. All authors read approved the final manuscript.

Corresponding authors

Correspondence to Dian-shuai Gao or Bao-Le Zhang.

Ethics declarations

Conflict of interest

The authors declared no potential conflict of interest.

Ethical Approval

Glioma tissues were obtained from the Affiliated Hospital of Xuzhou Medical University. All experimental protocols were approved by the Clinical Research Ethics Committee of the Affiliated Hospital of Xuzhou Medical University. All participates received and signed informed consent.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

Below is the link to the electronic supplementary material.

11064_2020_3140_MOESM1_ESM.tif

Supplementary Fig. 1. Prediction of Lysine ubiquitylation for CREB by UbPred using the CREB amino acid sequence.Supplementary file1 (TIF 7705 kb)

11064_2020_3140_MOESM2_ESM.tif

Supplementary Fig. 2. The original gel of complexes with the CUEDC2 antibody used for mass spectrometry.Supplementary file2 (TIF 4322 kb)

11064_2020_3140_MOESM3_ESM.tif

Supplementary Fig. 3. Transfected U251 cells were observed under the fluorescence microscope. The upper picture showed that transfected U251 cells were observed under a light microscope. The middle picture showed that transfected U251 cells were observed under the green fluorescence. Merge picture showed at the lowest picture. Supplementary file3 (TIF 14207 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Liu, XF., Tang, CX., Zhang, L. et al. Down-Regulated CUEDC2 Increases GDNF Expression by Stabilizing CREB Through Reducing Its Ubiquitination in Glioma. Neurochem Res 45, 2915–2925 (2020). https://doi.org/10.1007/s11064-020-03140-w

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11064-020-03140-w

Keywords

Search

Navigation