ATF5 increases cisplatin-induced apoptosis through up-regulation of Cyclin D3 transcription in HeLa cells

https://doi.org/10.1016/j.bbrc.2005.11.054Get rights and content

Abstract

ATF5 transcription factor plays an essential role in hematopoietic and glioma cell survival and neuronal cell differentiation. Here, we report for the first time the pro-apoptosis role of ATF5 and identify Cyclin D3 as an ATF5-targeted apoptosis-related gene. The ectopic expression of ATF5 in HeLa cells could markedly increase cisplatin-induced apoptosis and the cleavage of Caspase-3, and induce Cyclin D3 mRNA expression via cooperation with E2F1 transcription factor. Moreover, the interference of Cyclin D3 expression by transfection with Cyclin D3 RNAi could protect cells from ATF5-mediated apoptosis induced by cisplatin, indicating the contribution of Cyclin D3 in ATF5-mediated apoptosis. Taken together, these results suggest that ATF5 increases cisplatin-induced apoptosis through up-regulation of Cyclin D3 transcription, which elicits survival signals in HeLa cells.

Section snippets

Experimental procedures

Materials. Restriction enzymes, bovine calf serum, DMEM, Trizol reagent, LipofectAMINE reagent, and the expression vectors pcDNA3.0, pcDNA3.1 were purchased from Invitrogen. PGL3-Basic and pRL-CMV were from Promega. Hoechst33258, dimethyl sulfoxide, cisplatin, and etoposide were purchased from Sigma Chemical. The anti-ATF5 Ab was purchased from Abcam. The anti-Cyclin D3 Ab, anti-Cyclin D2 Ab, and anti-Cyclin D1 Ab were purchased from Pharmingen. The anti-E2F1 Ab and anti-GAPDH Ab were purchased

Ectopic expression of ATF5 increases apoptosis induced by Cisplatin in HeLa cells

To elucidate the role of ATF5 in the DNA damage agent-induced apoptosis, ATF5 expression vector or empty vector was transfected into HeLa cells (Fig. 1A). Next, we investigated the effect of ATF5 on apoptosis after cisplatin treatment for 12 h. As shown in Fig. 1B, ATF5 sensitized HeLa cells to cisplatin-induced apoptosis as indicated by fragmented and condensed nuclei, indicating the pro-apoptotic role of ATF5 in HeLa cells. This conclusion was further supported in Figs. 1C and D. The

Discussion

ATF5 is a member of a large family of transcription factors originally identified more than a decade ago [18]. Since then, additional ATFs that medicate cell apoptosis have been reported [19], [20], [21], [22]. In this report, we have described several observations that implicated the role of ATF5 in cisplatin-induced apoptosis in HeLa cells. (a) ATF5 was up-regulated in the apoptosis process induced by cisplatin, which has been widely used in cervical treatment [23]. (b) The ectopic expression

Acknowledgments

This work was supported by National Natural Scientific Foundation of China (30470442 and 30330320) and CNHLPP (2004BA711A19) and a grant from the Development of Science and Technology of Shanghai (02DJ14002).

References (30)

  • A. Blais et al.

    Regulation of the human cyclin-dependent kinase inhibitor p18INK4c by the transcription factors E2F1 and Sp1

    J. Biol. Chem.

    (2002)
  • H. Gunji et al.

    Induction of internucleosomal DNA fragmentation in human myeloid leukemia cells by 1-beta-d-arabinofuranosylcytosine

    Cancer Res.

    (1991)
  • S. Adachi et al.

    sc-Myc is necessary for DNA damage-induced apoptosis in the G(2) phase of the cell cycle

    Mol. Cell. Biol.

    (2001)
  • K.A. Lee et al.

    A cellular protein, activating transcription factor, activates transcription of multiple E1A-inducible adenovirus early promoters

    Proc. Natl. Acad. Sci. USA

    (1987)
  • E. Forgacs et al.

    The bZIP transcription factor ATFx binds human T-cell leukemia virus type 1 (HTLV-1) Tax and represses HTLV-1 long terminal repeat-mediated transcription

    J. Virol.

    (2005)
  • Cited by (17)

    • Nucleophosmin (NPM1/B23) interacts with activating transcription factor 5 (ATF5) protein and promotes proteasome- and caspase-dependent ATF5 degradation in hepatocellular carcinoma cells

      2012, Journal of Biological Chemistry
      Citation Excerpt :

      Re-expression of ATF5 in HCC inhibits cell proliferation and induces G2/M arrest (5). ATF5 has been shown to be involved in other cellular processes such as differentiation of neural progenitor cells (6–8), repression of cAMP-induced transcription in JEG3 choriocarcinoma cells and PC12 pheochromocytoma cells (6, 9), regulation of apoptosis (10–17), and response to various types of cellular stresses (18–20). ATF5 is known to subject to multilayered regulation that includes transcriptional regulation by EBF1 (21), translational regulation that is controlled by phosphorylated eIF2 (20, 23), and post-translational regulation that involves phosphorylation (2), acetylation (13), and Cdc34-dependent ubiquitin-mediated proteolysis (9, 24, 25).

    • Regulation of the human CHOP gene promoter by the stress response transcription factor ATF5 via the AARE1 site in human hepatoma HepG2 cells

      2010, Life Sciences
      Citation Excerpt :

      This suggested that up-regulation of ATF5 protein might be responsible for the up-regulation of CHOP expression. Wei et al. (2008) reported that ATF5 protein was up-regulated by cisplatin, and its overexpression increased cisplatin-induced apoptosis in HeLa cells (Wei et al. 2006). On the other hand, it has been reported that in an interleukin 3-dependent cell line, ATF5 suppresses apoptosis resulting from cytokine deprivation (Persengiev et al. 2002), promotes cell survival through transcriptional activation of Hsp27 in a rat embryonal cardiac myoblast cell line, H9c2 (Wang et al. 2007), and suppresses the transactivational activity of p53 and blocks the p53-dependent apoptosis induced by ionizing irradiation in a fibrosarcoma cell line (Nishioka et al. 2009).

    • Cdc34-mediated degradation of ATF5 is blocked by cisplatin

      2008, Journal of Biological Chemistry
      Citation Excerpt :

      Immunofluorescence and confocal assay were performed as previously described (22). Reverse Transcription-PCR and Co-immunoprecipitations—Reverse transcription-PCR was performed as described previously (5). Primers used for PCR were as follows: ATF5 sense, 5′-AAGTCGGCGGCTCTGAGGTA-3′, and ATF5 antisense, 5′-GGACTCTGCCCGTTCCTTCA-3′; P21 sense, 5′-GACACCACTGGAGGGTGACT-3′, and P21 antisense, 5′-GGCGTTTGGAGTGGTAGAAA-3′.

    • ATF5 promotes cell survival through transcriptional activation of Hsp27 in H9c2 cells

      2007, Cell Biology International
      Citation Excerpt :

      Taken together, our results showed that ATF5 could promote H9c2 cell survival during heat shock stress. This is not consistent with the report (Wei et al., 2006) that ATF5 was a pro-apoptotic factor in cisplatin treated HeLa cells. The pro-apoptotic or anti-apoptotic ability of ATF5 in response to different extracellular stimulation indicated that ATF5 may act as a negative regulator or/and positive regulator depending on promoter context.

    View all citing articles on Scopus
    View full text