Apoptosis Antagonizing Transcription Factor Research Articles

AATF (Apoptosis Antagonizing Transcription Factor)

Review on AATF (Apoptosis Antagonizing Transcription Factor), with data on DNA, on the protein encoded, and where the gene is implicated.

LXR-α genomics programmes neuronal death observed in Alzheimer’s disease

Keeping in view the fact that the most pathognomonic feature of Alzheimer's disease is the abnormal processing of neuronal cell membrane amyloid precursor protein accompanied by significantly elevated human serum and CSF levels of 24-hydroxycholesterol recognised widely as the specific endogenous ligand of Liver X receptor (LXR-α), the present study was addressed to explore the epigenomic-pathway (if any) that connects LXR-α activation with the genes recognised to be involved in the regulation of aberrant Abeta production leading to the generation of toxic and inflammatory mediators responsible for neuronal death. The results of such a study revealed that LXR-α activation by its specific endogenous or exogenous ligands within neuroblastoma cells resulted in the over-expression of PAR-4 gene accompanied by suppression of AATF gene through its inherent capacity to regulate genes coding for SREBP and NF-κB. Over-expression of PAR-4 gene was accompanied by aberrant Abeta production followed by ROS generation and subsequent death of neuroblastoma cells used in the present study as a cellular model for neurons. Further based upon these results, it was proposed that Abeta-induced heme oxygenase-1 can ensure cholesterol-oxidation to provide endogenous ligands for the sustained activation of neuronal LXR-α dependent epigenomic-pathway leading to neuronal death observed in Alzheimer's disease.

Functional genomics of tumor suppressor miR-196b in T-cell acute lymphoblastic leukemia

Huge data accumulated in last few years have shown that differential expression of candidate miRNAs in normal versus transformed cell provides important insights into the pathogenesis of cancer including leukemias. In our previous report, we have revealed that miR-196b was significantly down-regulated in both EB-3 cells as well as B-cell ALL (acute lymphoblastic leukemia) patients as compared to their respective controls. We have unambiguously proven that miR-196b restoration in EB-3 cells leads to significant down-regulation of c-myc and its effector genes, i.e., human telomerase reverse transcriptase (hTERT), B-cell lymphoma/leukemia-2 (Bcl-2), apoptosis antagonizing transcription factor (AATF), and qualifies for tumor suppressor function in B-cell ALL. Keeping in view these results, the present study was aimed at dissecting the role of miR-196b and other miRNAs present near/within the genomic regions involved in genetic translocations characteristic of ALL in T-cell ALL cell lines and patient samples. We have demonstrated significant down-regulation in the expression of miR-196b in MOLT-4 and T-cell ALL patients with respect to the respective control cells. Transfection experiments revealed that none of the six identified miRNAs were able to knock down the expression of c-myc gene. Interestingly, it was found that miR-196b loses its ability to down-regulate c-myc gene expression in T-cell ALL as a consequence of mutations in target 3'-untranslated region (3'-UTR) of the c-myc gene. Results of the present study revealed that miR-196b becomes non-functional in T-cell ALL as a consequence of mutations in 3'-UTR of c-myc gene in T-cell ALL cellular models.

A Novel Approach to Predict Protein-Protein Interactions Related to Alzheimers Disease Based on Complex Network

A protein domain interaction prediction program was complied using C(++) to predict whether a query protein could interact with other hub proteins. It is indicated that the proteins interacting with AATF (Apoptosis-Antagonizing Transcription Factor) possess a common conservative pattern: I-x(0,1)-E-x(2)-[A/E/N/T]-x-[E/K]. Our method is attempted to integrate the characteristics of protein interaction network with the simplest building blocks-conservative patterns.

Mutation analysis of the AATF gene in breast cancer families

BackgroundAbout 5-10% of breast cancer is due to inherited disease predisposition. Many previously identified susceptibility factors are involved in the maintenance of genomic integrity. AATF plays an important role in the regulation of gene transcription and cell proliferation. It induces apoptosis by associating with p53. The checkpoint kinases ATM/ATR and CHEK2 interact with and phosphorylate AATF, enhancing its accumulation and stability. Based on its biological function, and direct interaction with several known breast cancer risk factors, AATF is a good candidate gene for being involved in heritable cancer susceptibility.MethodsHere we have screened the entire coding region of AATF in affected index cases from 121 Finnish cancer families for germline defects, using conformation sensitive gel electrophoresis and direct sequencing.ResultsAltogether seven different sequence changes were observed, one missense variant and six intronic ones. Based on the in silico analyses of these sequence alterations, as well as their occurrence in cases and controls, none of them, however, were predicted to be pathogenic.ConclusionsTo our knowledge, this is the first study reporting the mutation screening of the AATF gene in familial breast cancer cases. No evidence for the association with breast cancer was observed.

AATF mediates an antiapoptotic effect of the unfolded protein response through transcriptional regulation of AKT1

Endoplasmic reticulum (ER) stress-mediated cell death plays an important role in the pathogenesis of chronic diseases including diabetes and neurodegeneration. Although pro-apoptotic programs activated by ER stress have been extensively studied, identification and characterization of anti-apoptotic programs that counteract ER stress is currently incomplete. Through the gene expression profiling of β-cells lacking WFS1, a causative gene for Wolfram syndrome, we have discovered a novel anti-apoptotic gene of the unfolded protein response (UPR), apoptosis antagonizing transcription factor (AATF). Here we study the regulation of AATF, identify its target genes, and determine the basis for its anti-apoptotic activities in response to ER stress. We show that AATF is induced by ER stress through the PERK-eIF2α pathway and transcriptionally activates the Akt1 gene through Stat3, which sustains Akt1 activation and promotes cell survival. Ectopic expression of AATF or a constitutively active form of AKT1 confers on cells resistance to ER stress-mediated cell death, whereas RNAi-mediated knockdown of AATF or AKT1 renders cells sensitive to ER stress. We also discovered positive crosstalk between the AATF and WFS1 signaling pathways. Thus, WFS1-deficiency or AATF-deficiency mediates a self-perpetuating cycle of cell death. Our results reveal a novel anti-apoptotic program relevant to treatment for diseases caused by ER stress-mediated cell death.

HIV-1 genome-encoded hiv1-mir-H1 impairs cellular responses to infection

The recent discovery of HIV-1 genome-encoded novel microRNA (miRNA; designated as hiv1-mir-H1) having ability to target selectively and specifically human cellular AATF gene, prompted us to explore the role of this miRNA in the regulation of genes involved in cellular apoptosis, proliferation and nucleic acid-based immune mechanism governed by miRNAs. Such a study revealed that this miRNA-induced knockdown of AATF gene, within normal human blood mononuclear cells, was responsible for the suppression of genes coding for Bcl-2, c-myc, Par-4 and Dicer. Further, hiv1-mir-H1 had the capacity to downregulate expression of cellular miR149 gene recognized to target Vpr gene encoded by HIV-1. Based upon these findings, we propose an "Epigenomic Pathway" through which hiv1-mir-H1 induced AATF gene knockdown within human mononuclear cells initiates their apoptosis.

Prediction of Butyrylcholinesterase Function Through Domain Analysis and Protein-Protein Interaction

The prediction of functions of proteins in living organisms can be made based on its domain analysis, proteinprotein interactions, sequence comparisons, structure comparisons, and phylogenetic tree analysis. Domain analysis and protein-protein interactions are important tools of bioinformatics based on which annotation of conserved domain footprints on protein sequences can be ascertained which could serve as the first step towards characterizing protein function(s) in silicon. In the present investigation, the functionality of butyrylcholinesterase (BChE) was ascertained by employing domain analysis and protein-protein interaction(s) tools. We collected the sequence of BChE, established the domains of this protein using NCBI conserved domain database (CDD), and identified proteins that interact with it (BChE). Using this data, we identified proteins with similar domain, proteins with which it interacted, and annotated the function of BChE via sequence similarity. These studies revealed that BChE interacts with proteins: ubiquitin-conjugating enzyme, apoptosisantagonizing transcription factor, paraoxonase/arylesterase, pyruvate dehydrogenase protein X component, proteasome activator complex subunit 3, 4-trimethyl-aminobutyraldehyde dehydrogenase, and vacuolar proton translocating ATPase whose functions are known. Based on this information, it is predicted that BChE may have actions similar to those with which it interacts. Thus, by using domain analysis and protein-protein interaction(s) tools it may be possible to identify the functions of proteins whose action is unknown by annotating the conserved domain amino acid sequence of proteins whose function is known with those of the unknown.

ZIP kinase plays a crucial role in androgen receptor-mediated transcription

The androgen receptor (AR) is a ligand-dependent transcription factor that plays a crucial role in the development and homeostasis of the prostate and in prostate cancer. The transcriptional activity of AR is mediated by interaction with multiple co-activators, which serve in chromatin modification or remodeling, or provide a link between specific and general transcription factors. We have identified zipper interacting protein (ZIP) kinase as a novel transcriptional co-activator of the AR. ZIP kinase enhanced expression of AR-responsive promotor/luciferase reporter constructs in a hormone- and kinase-dependent manner. Similar results were obtained for glucocorticoid receptor but not for progesterone receptor and estrogen receptor. Following hormone treatment, AR and ZIP kinase formed physical complexes and associated with the promoter and enhancer of the prostate-specific antigen gene, as revealed by chromatin immunoprecipitation. Strikingly, depletion of ZIP kinase by siRNA led to significant reduction of AR-mediated transactivation. The interaction of ZIP kinase with AR seems to be mediated in part by apoptosis antagonizing transcription factor and in part by direct binding. Interestingly, AR was not phosphorylated by ZIP kinase in vitro, suggesting that it phosphorylates other co-activators or chromatin proteins.

NRAGE associates with the anti-apoptotic factor Che-1 and regulates its degradation to induce cell death

Neurotrophin receptor-interacting MAGE homolog (NRAGE) has been recently identified as a cell-death inducer, involved in molecular events driving cells through apoptotic networks during neuronal development. Recently, we have focused on the functional role of Che-1, also known as apoptosis-antagonizing transcription factor (AATF), a protein involved in cell cycle control and gene transcription. Increasing evidence suggests that Che-1 is involved in apoptotic signalling in neural tissues. In cortical neurons Che-1 exhibits an anti-apoptotic activity, protecting cells from neuronal damage induced by amyloid beta-peptide. Here, we report that Che-1 interacts with NRAGE and that an EGFP-NRAGE fusion protein inhibits nuclear localization of Che-1, by sequestering it within the cytoplasmic compartment. Furthermore, NRAGE overexpression downregulates endogenous Che-1 by targeting it for proteasome-dependent degradation. Finally, we propose that Che-1 is a functional antagonist of NRAGE, because its overexpression completely reverts NRAGE-induced cell-death.

Apoptosis Antagonizing Transcription Factor Protects Renal Tubule Cells against Oxidative Damage and Apoptosis Induced by Ischemia-Reperfusion

Apoptosis antagonizing transcription factor (AATF) is a leucine zipper domain-containing protein that has antiapoptotic properties. AATF is expressed in several organs and tissues, including the kidney. AATF may participate in inhibition of proapoptotic pathways and/or activation of antiapoptotic pathways. Ischemia/reperfusion-induced renal injury (IRI) is clinically important because it typically damages renal tubular epithelial cells and glomerular cells and is the most common cause of acute renal failure. It now is reported that AATF is expressed in human kidney proximal tubule (HK-2) cells and in mouse primary renal tubule epithelial cells. Levels of AATF expression were altered significantly in these cells in a well-established in vitro model of renal IRI. In transfected HK-2 cells, RNA interference-mediated silencing of AATF exacerbated whereas overexpression of the full-length AATF ameliorated mitochondrial dysfunction, accumulation of superoxide and peroxynitrite, lipid peroxidation, caspase-3 activation, and apoptotic death that were induced by IRI. In primary renal tubule epithelial cells, overexpression of AATF mediated by recombinant adeno-associated virus (AAV) vectors resulted in significant antiapoptotic activity, whereas knockdown of AATF by small interference RNA led to exacerbated cell death after IRI. These results identify AATF as a novel cytoprotective factor against oxidative and apoptotic damage in renal tubular cells. AATF may represent a potential candidate for therapeutic application in IRI.

Functional characterization of AATF transcriptome in human leukemic cells

The study, addressed to explore the transcriptional expression and regulation of Apoptosis-antagonizing transcription factor (AATF) gene within various types of human leukemic cell lines, revealed that AATF gene was overexpressed ubiquitously in all the leukemic cell lines studied and this upregulation was accompanied by c-myc gene overamplification in these cells. Downregulation of AATF gene transcription within leukemic cells not only resulted in the downregulation of c-myc gene and vice-versa but also contributed to apoptosis leading to cell death. Further, the link between AATF expression and leukemic cellular apoptosis involved PI3K/Akt pathway. Based on these results we propose that AATF gene may be of crucial importance in maintaining the leukemic state of a cell compartment through its ability to initiate cell proliferation coupled with repression of cellular apoptosis.

TSG101 Interacts with Apoptosis-antagonizing Transcription Factor and Enhances Androgen Receptor-mediated Transcription by Promoting Its Monoubiquitination

Apoptosis-antagonizing transcription factor (AATF), also termed Che-1, was identified as interacting protein of Dlk/ZIP kinase and RNA polymerase II, respectively. Che-1 has additionally been shown to bind Rb, thereby activating transcription factor E2F and promoting cell cycle progression. Moreover, AATF enhances steroid receptor-mediated transactivation in a hormone- and dose-dependent manner (Leister, P., Burgdorf, S., and Scheidtmann, K. H., (2003) Signal Transduction 3, 18-25). These data suggest that AATF exerts its functions through interaction with different transcription factors. In search of novel interaction partners of AATF, we identified the tumor susceptibility gene product TSG101, which had also been recognized as a co-regulator of nuclear hormone receptors. Interestingly, TSG101 and AATF functioned as cooperative coactivators in androgen receptor-mediated transcription. Because TSG101 was also shown to play a role in regulation of ubiquitin conjugation, we asked whether its coactivating function might be linked to ubiquitination. Indeed, TSG101 enhanced monoubiquitination of the androgen receptor in a ligand-dependent manner, and this correlated with enhanced transactivating capacity. Furthermore, a dominant-negative mutant of ubiquitin preventing polyubiquitination also stimulated androgen receptor-mediated transcription, which in this case could not be enhanced by TSG101. We propose that TSG101 activates androgen receptor-induced transcription by transient stabilization of the monoubiquitinated state, thus revealing a novel regulatory mechanism for nuclear receptors.

AATF protects neural cells against oxidative damage induced by amyloid β-peptide

Extensive loss of neurons and synapses in vulnerable regions of the brain is one of the most important pathological features of Alzheimer's disease (AD). Increased oxidative stress has been shown to contribute to the neurodegenerative process in AD. Aggregation of amyloid β-peptide (Aβ) in amyloid plaques is one of the defining features of Alzheimer's disease. Indeed, Aβ has been shown to induce oxidative stress and apoptosis in many in vivo and in vitro models of AD. We now report that AATF (apoptosis-antagonizing transcription factor), a leucine zipper protein initially identified as an interaction partner of DAP like kinase (Dlk, a member of the pro-apoptotic Death-Associated Protein kinase family), is expressed in cortical neurons and in neural PC12 cells. Aβ induces alterations in AATF expression in cortical neurons. Inhibition of AATF induction sensitizes neurons to Aβ toxicity. Overexpression of AATF suppressed superoxide production, inhibited peroxynitrite formation and membrane lipid peroxidation, and protected against Aβ-induced apoptosis in PC12 cells. These results suggest that AATF is a novel neuroprotective factor and it may protect against Aβ-induced apoptosis through its effects on suppressing the production of reactive oxygen species (ROS). AATF may therefore represent a potential candidate for therapeutic intervention of neurodegeneration in both sporadic and familial forms of AD.

AATF Inhibits Aberrant Production of Amyloid β Peptide 1-42 by Interacting Directly with Par-4

Aggregation of the neurotoxic amyloid beta peptide 1-42 (Abeta-(1-42)) in the brain is considered to be an early event in the pathogenesis of Alzheimer's disease (AD). Par-4 (prostate apoptosis response-4) is a leucine zipper protein that is pro-apoptotic and associated with neuronal degeneration in AD. Overexpression of Par-4 significantly increased production of Abeta-(1-42) after initiation of apoptotic cascades, indicating factors regulating apoptotic pathways may also affect processing of beta-amyloid precursor protein (APP). AATF (apoptosis-antagonizing transcription factor) was recently identified as an interaction partner of DAP-like kinase (Dlk), a member of the DAP (death-associated protein) kinase family. AATF antagonizes apoptosis induced by Par-4, suggesting that AATF might directly or indirectly participate in regulation of Par-4 activity. We now report that AATF colocalizes with Par-4 in both cytoplasmic and nuclear compartments, and it interacts directly and selectively with Par-4 via the leucine zipper domain in neural cells. Par-4 induced an aberrant production and secretion of Abeta in neuroblastoma IMR-32 cells after apoptotic cascades are initiated. Co-expression of AATF completely blocked aberrant production and secretion of Abeta-(1-42) induced by Par-4, and AATF/Par-4 complex formation was essential for the inhibitory effect of AATF on aberrant Abeta secretion. These results indicate that AATF is an endogenous antagonist of Par-4 activity and an effective inhibitor of aberrant Abeta production and secretion under apoptotic conditions.

Apoptosis antagonizing transcription factor AATF is a novel coactivator of nuclear hormone receptors

AbstractApoptosis antagonizing transcription factor AATF was identified as interaction partner of Dlk/ZIP kinase and shown to exhibit transactivation activity in Gal4‐based reporter gene expression assays. The human ortholog Che‐1 was independently isolated as interacting protein of RNA polymerase II suggesting that AATF might be a transcriptional adapter or mediator which serves to link transcriptional activators to the general transcription machinery. Consistent with this assumption, AATF contains several interaction motifs including a leucine zipper and three putative binding motifs for nuclear hormone receptors. In this study, we investigated a possible functional relationship of AATF to nuclear receptors. Indeed, AATF enhanced transactivation by several steroid hormone receptors including androgen, estrogen and glucocorticoid receptors in a hormone‐dependent manner. This enhancing effect was further increased by p300 but not by a mutant of p300 lacking acetyl transferase activity. AATF bound directly to the receptors in vitro. Co‐activation was impaired with AATF mutants bearing mutations in the LXXLL NR binding motifs. Our data suggest that AATF is a novel coactivator of nuclear hormone receptors.

Regulation of microglial expression of integrins by poly (ADP-ribose) polymerase-1

The molecular mechanism of DNA damage induced by hydroquinone (HQ) remains unclear. Poly(ADP-ribose) polymerase-1 (PARP-1) usually works as a DNA damage sensor, and hence, it is possible that PARP-1 is involved in the DNA damage response induced by HQ. In TK6 cells treated with HQ, PARP activity as well as the expression of apoptosis antagonizing transcription factor (AATF), PARP-1, and phosphorylated H2AX (γ-H2AX) were maximum at 0.5 h, 6 h, 3 h, and 3 h, respectively. To explore the detailed mechanisms underlying the prompt DNA repair reaction, the above indicators were investigated in PARP-1-silenced cells. PARP activity and expression of AATF and PARP-1 decreased to 36%, 32%, and 33%, respectively, in the cells; however, γ-H2AX expression increased to 265%. Co-immunoprecipitation (co-IP) assays were employed to determine whether PARP-1 and AATF formed protein complexes. The interaction between these proteins together with the results from IP assays and confocal microscopy indicated that poly(ADP-ribosyl)ation (PARylation) regulated AATF expression. In conclusion, PARP-1 was involved in the DNA damage repair induced by HQ via increasing the accumulation of AATF through PARylation.

AATF, a novel transcription factor that interacts with Dlk/ZIP kinase and interferes with apoptosis

Dlk, also known as ZIP kinase, is a serine/threonine kinase that is tightly associated with nuclear structures. Under certain conditions, which require cytoplasmic localization, Dlk can induce apoptosis. In search for interaction partners that might serve as regulators or targets of this kinase we identified apoptosis antagonizing transcription factor (AATF), a nuclear phosphoprotein of 523 amino acids. The 1.8 kb mRNA seems to be ubiquitously expressed. AATF contains an extremely acidic domain and a putative leucine zipper characteristic of transcription factors. Indeed, a Gal4-BD-AATF fusion protein exhibited strong transactivation activity. Interestingly, AATF interfered with Dlk-induced apoptosis.