ATF3 induction following DNA damage is regulated by distinct signaling pathways and over-expression of ATF3 protein suppresses cells growth
Authors: Shunqian Jin
Baskaran Rajasekaran
Sally A Amundson
Tong Tong
Xianxing Li
Hongcheng Zhao
Albert J Fornace Jr
Feiyue Fan
Kimberly L Petrik
Xiaocheng Zhu
Wenhong Fan
Qimin Zhan
Lucia Mazzacurati
Journal: Oncogene (2002)
Discipline: MOLECULAR BIOLOGY & GENETICS
Keywords: Mammalian cells; genotoxic stress; DNA damage; cellular responses; gene induction
Affiliations: Department of Radiation Oncology, Cancer Institute, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania PA 15213, USA.

Abstract:  Mammalian cells have a remarkable diverse repertoire of response to genotoxic stress that damage DNA. Cellular responses to DNA damaging agents will initially exhibit gene induction, which is regulated by complex mechanism(s) and probably involves multiple signaling pathways. In this paper, we demonstrate that induction of ATF3 protein, a member of the ATF/CREB family of transcription factors, by ionizing radiation (IR) requires normal cellular p53 function. In contrast, induction of ATF3 after UV radiation (UV) or Methyl methanesulphonate (MMS) is independent of p53 status. Induction of ATF3 by DNA damage is rapid, transient, and through a transcriptional mechanism. The ATF3 promoter is induced by UV and MMS, but not by IR. In addition, ATF3 promoter can be activated by MEKK1, an upstream activator of the ERK and JNK kinase pathway, but not induced following p53 expression. Those results indicate that regulation of ATF3 induction after DNA damage utilizes both the p53-dependent and -independent pathways, and may also involve MAP kinase signaling pathways. Using the tetracycline-inducible system (tet-off), we have found that over-expression of ATF3 protein moderately suppresses cell growth. Interestingly, over-expression of ATF3 protein is able to slow down progression of cells from G1 to S phase, indicating that ATF3 protein might play a negative role in the control of cell cycle progression. 

Retracted: No

PubPeer comments:
 There might be some problems with the primers in this paper. Firstly, primers for Gadd45 amplification may be invalid according the In-Silico result (see figure below). Secondly, primers for beta-actin were simply copied the primers for AFT3. Finally, primers for Oct1 was not provide. ![file] , 
 Immunoblotting of Actin in Figure 5A was reappeared in another paper published by the same team (DOI: 10.1038 sj.onc.1205896). See figure below. ![file] , 
 Dear Cicindela Chrysippe, Thank you for your interest of our study and pointing these out. In this study, we focused on the regulation of ATF3 upon DNA damage and actually did not include Gadd45 and Oct1 in this study. Thus, these were clerical error during preparing the manuscript. Additionally, the reverse primer of u03b2-actin was a mistake by copy error. We have contacted with the editor office of Oncogene for a corrigendum. Again thank you for your valuable comment. #2. Immunoblotting of Actin in Figure 5A was reappeared in another paper published by the same team (DOI: 10.1038 sj.onc.1205896). Dear Cicindela Chrysippe, Thank you for your interest of our study and carefully reading our paper. By checking back our original data, we confirm that the internal control Actin in this study was reused by mistake. Here we provide the corrected Figure 5A as below. Now we have contacted with the editor office of Oncogene for a corrigendum. Again many thanks for your comment. Corrected Figure 5Auff1a ![file] Figure 5 Cell growth suppression following expression of ATF3 protein. (A) Hela-ATF3-inducible cell line was established as described in Materials and methods. Cells were placed in 100 mm dishes at a density of 4u00d7105 and grown in DMEM medium containing tetracycline at a concentration of 2 u03bcg ml. After withdrawal of tetracycline, cells were collected at the indicated time points for preparation of cellular protein. One hundred u03bcg of whole cell protein was used for immunoblotting analysis with anti-ATF3 antibody. As a loading control, anti-actin antibody was included. ,