DOCSLIB.ORG

Nuclear Localization of DP and E2F Transcription Factors by Heterodimeric Partners and Retinoblastoma Protein Family Members

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Nuclear Localization of DP and E2F Transcription Factors by Heterodimeric Partners and Retinoblastoma Protein Family Members Journal of Cell Science 109, 1717-1726 (1996) 1717 Printed in Great Britain © The Company of Biologists Limited 1996 JCS7086 Nuclear localization of DP and E2F transcription factors by heterodimeric partners and retinoblastoma protein family members Junji Magae1, Chin-Lee Wu2, Sharon Illenye1, Ed Harlow2 and Nicholas H. Heintz1,* 1Department of Pathology, University of Vermont, Burlington VT 05405, USA 2Massachusetts General Hospital Cancer Center, Charlestown MA 02129, USA *Author for correspondence SUMMARY E2F is a family of transcription factors implicated in the showed that regions of E2F-1 and DP-1 that are required regulation of genes required for progression through G1 for stable association of the two proteins were also required and entry into the S phase. The transcriptionally active for nuclear localization of DP-1. Unlike E2F-1, -2, and -3, forms of E2F are heterodimers composed of one polypep- E2F-4 did not accumulate in the nucleus unless it was coex- tide encoded by the E2F gene family and one polypeptide pressed with DP-2. p107 and p130, but not pRb, stimulated encoded by the DP gene family. The transcriptional activity nuclear localization of E2F-4, either alone or in combina- of E2F/DP heterodimers is influenced by association with tion with DP-2. These results indicate that DP proteins the members of the retinoblastoma tumor suppressor preferentially associate with specific E2F partners, and protein family (pRb, p107, and p130). Here the intracellu- suggest that the ability of specific E2F/DP heterodimers to lar distribution of E2F and DP proteins was investigated in localize in the nucleus contributes to the regulation of E2F transiently transfected Chinese hamster and human cells. activity. In transfected cells, DP-1 did not accumulate in the nucleus unless it was coexpressed with the heterodimeric partners E2F-1, E2F-2, or E2F-3. Domain mapping experiments Key words: Gene expression, Cell cycle, Protein trafficking INTRODUCTION by cyclin-dependent kinases inhibits E2F DNA binding activity and E2F-dependent transcription (Krek et al., 1994; First identified as a cellular factor required for the transcrip- Dynlacht et al., 1994; Xu et al., 1994). Disruption of pRb/E2F tional activation of the adenovirus E2 gene promoter (reviewed complexes by viral oncoproteins is essential for cell transfor- by Moran, 1993; Nevins, 1992), E2F has been implicated in mation by DNA tumor viruses, and mutations in pRb that the periodic regulation of cellular genes required for transition disrupt interactions with E2F are implicated in both sporadic through G1 and entry into the S phase (reviewed by Farnham and inherited forms of human cancer (reviewed by Dyson, et al., 1993; Horowitz, 1993; La Thangue, 1994; Lam and 1994; Ewen, 1994; Lam and La Thangue, 1994; Wiman, 1993). LaThangue, 1994; Muller, 1995; Wiman, 1993). The evidence Moreover, deregulated expression of E2F genes leads to mor- that E2F plays a central role in regulating progression through phological changes (Logan et al., 1994; Shan and Lee, 1994; the cell cycle is compelling. E2F forms higher order complexes Xu et al., 1995), entry into the S phase (Almasan et al., 1995; with a number of proteins that regulate progression through the Johnson et al., 1993; Kowalik et al., 1995; Sardet et al., 1995; cell cycle, including the products of the retinoblastoma tumor Shan and Lee, 1994), and cell transformation (Singh et al., suppressor gene family (i.e. pRb, p107 and p130) (Chellappan 1994; Xu et al., 1995; Yang and Sladek, 1995), and, under et al., 1991; Chittenden et al., 1991; Cao et al., 1992; Cobrinik some conditions, apoptosis (Qin et al., 1994; Shan and Lee, et al., 1993; Dyson et al., 1993; Huang et al., 1993; Krek et al., 1994; Wu and Levine, 1994; Kowalik et al., 1995). 1993; Lees et al., 1993; Fagan et al., 1994; Kim et al., 1994; The regulation of E2F by pRb family members and cyclin- Qin et al., 1995) and several cyclin-dependent kinases (Mudryj dependent kinases is exceedingly complex. Although the asso- et al., 1991; Devoto et al., 1992; Lees et al., 1992; Pagano et ciation of regulatory factors with E2F is tightly regulated al., 1992; Ewen et al., 1993; Kato et al., 1993; Dynlacht et al., during the cell cycle (Lees et al., 1992; Pagano et al., 1992; 1994). Association of pRb family members with E2F sup- Shirodkar et al., 1992; Schwarz et al., 1993; Chittenden et al., presses E2F-dependent transcription (Hiebert et al., 1992; 1993; Cobrinik et al., 1993; Hijmans et al., 1995), the sequence Flemington et al., 1993; Helin et al., 1993a; Hiebert, 1993; of these interactions during the cell cycle does not appear to Zamanian and La Thangue, 1993; Qin et al., 1995; Smith and be the same for all cell types. In addition, E2F participates in Nevins, 1995) and cell growth (Hiebert, 1993; Zhu et al., 1993; the transcriptional regulation of genes encoding members of Vairo et al., 1995; Zhu et al., 1995a). Phosphorylation of E2F the E2F/pRb regulatory loop, including the pRb, p107 and 1718 J. Magae and others E2F-1 genes (Hsiao et al., 1994; Johnson et al., 1994b; al., 1981), and the human cell lines U2OS and HeLa were cultured in Neuman et al., 1994; Zhu et al., 1995b). a 5% CO2 atmosphere at 37°C using D-MEM supplemented with 5% The transcriptionally active forms of E2F are a collection of FBS (Gibco). For DNA transfections, 1×106 CHOC 400 cells were heterodimeric protein complexes (Girling et al., 1993; Helin et plated in 85 mm culture dishes in 10 ml DMEM with 5% fetal bovine serum (FBS), incubated overnight, changed into fresh culture al., 1993b; Huber et al., 1993; Wu et al., 1995; Zhang and Chel- µ lappan, 1995), each composed of one E2F protein family subunit medium, and 6 hours later were transfected with 24 g of DNA using the calcium phosphate precipitation method as described previously and one DP protein family subunit. (Here we use E2F to refer (Helin et al., 1993b; Wu et al., 1995). The E2F and DP pCMV to the collection of heterodimeric complexes formed by the asso- expression vectors (Helin et al., 1993b; Wu et al., 1995) were used at ciation of specific E2F and DP family members, with the indi- 8 µg/culture dish with pBSK as carrier DNA. After incubation vidual components of each complex identified by the name of overnight, cells were washed twice with phosphate buffered saline the cloned cDNA.) To date cDNA clones for seven members of (PBS, pH 7.5), incubated in fresh medium for 4 hours, trypsinized, the mammalian E2F and DP gene families have been isolated. plated on coverslips, and incubated for an additional 20 hours in Interaction with pRb was used to clone E2F-1 (Helin et al., 1992; culture medium. Kaelin et al., 1992; Shan et al., 1992; Li et al., 1994), and Immunostaining homology with E2F-1 was then used to isolate E2F-2, E2F-3, and E2F-4 (Ivey-Hoyle et al., 1993; Lees et al., 1993; Ginsberg Cells on coverslips were washed with PBS, fixed with 4% paraformaldehyde in PBS for 15 minutes, permeabilized with 0.2% et al., 1994). E2F-4 was also cloned by virtue of its specific inter- Triton-X in PBS for 15 minutes, and blocked in PBS containing 0.1% action with p107 (Beijersbergen et al., 1994), and both E2F-4 azide, 0.1% Tween-20 and 2% FBS for 60 minutes. Coverslips were and E2F-5 were isolated in a screen for proteins that interact with then incubated with primary murine antibodies at a 1:5 dilution of p130 (Hijmans et al., 1995; Sardet et al., 1995). DP-1 was cloned culture supernatant, or 1:200 dilution of polyclonal mouse serum, in as a protein component of DRTF, a developmentally-regulated blocking buffer for 60 minutes. The coverslips were then washed three E2F-like activity from mouse embryonal F9 cells (Girling et al., times in Tris-buffered saline (pH 7.5) containing 0.1% Tween-20 1993). Recently, homology with DP-1 was used to isolate DP- (TBS-T), and incubated with FITC-conjugated anti-mouse 2, a second member of the mammalian DP protein family (Wu immunoglobulin diluted 1:100 in blocking buffer for 60 minutes. After washing three times in TBS-T, coverslips were incubated in 1 et al., 1995; Zhang and Chellappan, 1995). Comparison of the µ amino acid sequences of mammalian E2F family members g/ml propidium iodide (PI) in TBS-T for 60 minutes at 50°C, washed with TBS-T, mounted and photographed with an Olympus BX50 suggests E2F-4 and E2F-5 are highly related to one another, and microscope using a WIBA filter for FITC, WG filter for PI, and MT represent a subclass of factors distinct from E2F-1, -2 and -3 filter for PI/FITC. (Sardet et al., 1995). Both DP-1 and DP-2 dimerize with several E2F family members to form an array of transcriptionally active Antibodies forms of E2F/DP heterodimers (Bandara et al., 1993; Girling et The primary antibodies were monoclonal antibody KH20 for E2F-1 al., 1993; Helin et al., 1993b; Huber et al., 1993; Wu et al., 1995; (Helin et al., 1993b), monoclonal antibody WTH10 for DP-1 (Wu et Zhang and Chellappan, 1995). al., 1995), polyclonal antibodies for E2F-2, E2F-3, or E2F-4, and The function of individual E2F/DP heterodimers is not either a polyclonal antibody against DP-2 or monoclonal antibody known. While all E2F/DP complexes stimulate transcription of 12CA5 for HA-tagged DP-2 and HA-tagged E2F-3. Polyclonal anti- reporter genes from consensus E2F binding sites, different E2F bodies for E2F-2, E2F-3, E2F-4 and DP-2 were made against His- tagged full length proteins in mice and characterized as described (Wu complexes may regulate E2F-dependent cellular promoters et al., 1995).
Load more

Recommended publications
  • Retinoblastoma Protein Positively Regulates Terminal Adipocyte Differentiation Through Direct Interaction with C/Ebps
    Downloaded from genesdev.cshlp.org on October 3, 2021 - Published by Cold Spring Harbor Laboratory Press Retinoblastoma protein positively regulates terminal adipocyte differentiation through direct interaction with C/EBPs Phang-Lang Chen, Daniel J. Riley, Yumay Chen, and Wen-Hwa Lee l Center for Molecular Medicine, Institute of Biotechnology, The University of Texas Health Science Center at San Antonio, San Antonio, Texas 78245 USA To define a mechanism by which retinoblastoma protein (Rb) functions in cellular differentiation, we studied primary fibroblasts from the lung buds of wild-type (RB + / +) and null-mutant (RB-/-) mouse embryos. In culture, the RB +/+ fibroblasts differentiated into fat-storing cells, either spontaneously or in response to hormonal induction; otherwise syngenic RB -/- fibroblasts cultured in identical conditions did not. Ectopic expression of normal Rb, but not Rb with a single point mutation, enabled RB-/- fibroblasts to differentiate into adipocytes. Rb appears in murine fibroblasts to activate CCAAT/enhancer-binding proteins (C/EBPs), a family of transcription factors crucial for adipocyte differentiation. Physical interaction between Rb and C/EBPs was demonstrated by reciprocal coimmunoprecipitation, but occurred only in differentiating cells. Wild-type Rb also enhanced the binding of C/EBP to cognate DNA sequences in vitro and the transact[vat[on of a C/EBPl$-responsive promoter in cells. Taken together, these observations establish a direct and positive role for Rb in terminal differentiation. Such a role contrasts with the function of Rb in arresting cell cycle progression in G1 by negative regulation of other transcription factors like E2F-1. [Key Words: C/EBP; transcription factors; cell cycle; adipocyte differentiation; tumor suppressor protein] Received July 18, 1996; revised version accepted September 5, 1996.
    [Show full text]
  • TP63 and TP73 in Cancer, an Unresolved В€Œfamily∕ Puzzle Of
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector FEBS Letters 588 (2014) 2590–2599 journal homepage: www.FEBSLetters.org Review TP63 and TP73 in cancer, an unresolved ‘‘family’’ puzzle of complexity, redundancy and hierarchy Antonio Costanzo a, Natalia Pediconi b,c, Alessandra Narcisi a, Francesca Guerrieri c,d, Laura Belloni c,d, ⇑ Francesca Fausti a, Elisabetta Botti a, Massimo Levrero c,d, a Dermatology Unit, Department of Neuroscience, Mental Health and Sensory Organs (NESMOS), Sapienza University of Rome, Italy b Laboratory of Molecular Oncology, Department of Molecular Medicine, Sapienza University of Rome, Italy c Center for Life Nanosciences (CNLS) – IIT/Sapienza, Rome, Italy d Laboratory of Gene Expression, Department of Internal Medicine (DMISM), Sapienza University of Rome, Italy article info abstract Article history: TP53 belongs to a small gene family that includes, in mammals, two additional paralogs, TP63 and Received 19 May 2014 TP73. The p63 and p73 proteins are structurally and functionally similar to p53 and their activity as Revised 16 June 2014 transcription factors is regulated by a wide repertoire of shared and unique post-translational mod- Accepted 16 June 2014 ifications and interactions with regulatory cofactors. p63 and p73 have important functions in Available online 28 June 2014 embryonic development and differentiation but are also involved in tumor suppression. The biology Edited by Shairaz Baksh, Giovanni Blandino of p63 and p73 is complex since both TP63 and TP73 genes are transcribed into a variety of different and Wilhelm Just isoforms that give rise to proteins with antagonistic properties, the TA-isoforms that act as tumor- suppressors and DN-isoforms that behave as proto-oncogenes.
    [Show full text]
  • RB1 and P53 at the Crossroad of EMT and Triple Negative Breast Cancer
    PERSPECTIVE PERSPECTIVE Cell Cycle 10:10, 1-8; May 15, 2011; © 2011 Landes Bioscience RB1 and p53 at the crossroad of EMT and triple negative breast cancer Zhe Jiang,1 Robert Jones,1 Jeff C. Liu,1 Tao Deng,1 Tyler Robinson,1 Philip E.D. Chung,1 Sharon Wang,1 Jason I. Herschkowitz,2 Sean E. Egan,3 Charles M. Perou4 and Eldad Zacksenhaus1,* 1Division of Cell and Molecular Biology; Toronto General Research Institute; University Health Network; Toronto, Ontario, Canada; 2Department of Molecular and Cellular Biology; Baylor College of Medicine; Houston, TX USA; 3Program in Developmental and Stem Cell Biology; The Hospital for Sick Children; Department of Molecular Genetics; University of Toronto; Toronto, Ontario, Canada; 4Lineberger Comprehensive Cancer Center; Department of Genetics and Pathology; University of North Carolina at Chapel Hill; Chapel Hill, NC USA riple negative breast cancer (TNBC) NEU-positive and Triple Negative (TN) Tis a heterogeneous disease that tumors, the latter of which do not express includes Basal-like and Claudin-low hormone receptors or HER2.1-7 TNBC tumors. The Claudin-low tumors are affects 15–30% of patients. By IHC it can enriched for features associated with be further divided into Basal-like breast epithelial-to-mesenchymal transition cancer and non-basal tumors, some of (EMT) and possibly for tumor initiating which exhibit features of EMT.8 Basal-like cells. Primary TNBCs respond relatively BCs express the basal cytokeratins (CK) well to conventional chemotherapy; CK5/6, CK14, CK17, and/or epidermal © 2011 Landes Bioscience. Landes ©2011 however, metastatic disease is virtually growth factor receptor (EGFR), whereas incurable.
    [Show full text]
  • A Thyroid Hormone Receptor Coactivator Negatively Regulated by the Retinoblastoma Protein
    Proc. Natl. Acad. Sci. USA Vol. 94, pp. 9040–9045, August 1997 Biochemistry A thyroid hormone receptor coactivator negatively regulated by the retinoblastoma protein KAI-HSUAN CHANG*†,YUMAY CHEN*†,TUNG-TI CHEN*†,WEN-HAI CHOU*†,PHANG-LANG CHEN*, YEN-YING MA‡,TERESA L. YANG-FENG‡,XIAOHUA LENG§,MING-JER TSAI§,BERT W. O’MALLEY§, AND WEN-HWA LEE*¶ *Department of Molecular Medicine and Institute of Biotechnology, University of Texas Health Science Center at San Antonio, 15355 Lambda Drive, San Antonio, TX 78245; ‡Department of Genetics, Obstetrics, and Gynecology, Yale University School of Medicine, New Haven, CT 06510; and §Department of Cell Biology, Baylor College of Medicine, Houston, TX 77030 Contributed by Bert W. O’Malley, June 9, 1997 ABSTRACT The retinoblastoma protein (Rb) plays a E2F-1, a transcription factor important for the expression of critical role in cell proliferation, differentiation, and devel- several genes involved in cell cycle progression from G1 to S opment. To decipher the mechanism of Rb function at the (18). Rb inhibits E2F-1 activity by blocking its transactivation molecular level, we have systematically characterized a num- region (19–21). In contrast, Rb has been shown to have the ber of Rb-interacting proteins, among which is the clone C5 ability to increase the transactivating activity of the members described here, which encodes a protein of 1,978 amino acids of the CCAATyenhancer binding protein (CyEBP) family, and with an estimated molecular mass of 230 kDa. The corre- to be required for CyEBPs-dependent adipocyte and mono- sponding gene was assigned to chromosome 14q31, the same cytes differentiation (16–17).
    [Show full text]
  • AP-1 in Cell Proliferation and Survival
    Oncogene (2001) 20, 2390 ± 2400 ã 2001 Nature Publishing Group All rights reserved 0950 ± 9232/01 $15.00 www.nature.com/onc AP-1 in cell proliferation and survival Eitan Shaulian1 and Michael Karin*,1 1Laboratory of Gene Regulation and Signal Transduction, Department of Pharmacology, University of California San Diego, 9500 Gilman Drive, La Jolla, California, CA 92093-0636, USA A plethora of physiological and pathological stimuli extensively discussed previously (Angel and Karin, induce and activate a group of DNA binding proteins 1991; Karin, 1995). that form AP-1 dimers. These proteins include the Jun, The mammalian AP-1 proteins are homodimers and Fos and ATF subgroups of transcription factors. Recent heterodimers composed of basic region-leucine zipper studies using cells and mice de®cient in individual AP-1 (bZIP) proteins that belong to the Jun (c-Jun, JunB proteins have begun to shed light on their physiological and JunD), Fos (c-Fos, FosB, Fra-1 and Fra-2), Jun functions in the control of cell proliferation, neoplastic dimerization partners (JDP1 and JDP2) and the closely transformation and apoptosis. Above all such studies related activating transcription factors (ATF2, LRF1/ have identi®ed some of the target genes that mediate the ATF3 and B-ATF) subfamilies (reviewed by (Angel eects of AP-1 proteins on cell proliferation and death. and Karin, 1991; Aronheim et al., 1997; Karin et al., There is evidence that AP-1 proteins, mostly those that 1997; Liebermann et al., 1998; Wisdom, 1999). In belong to the Jun group, control cell life and death addition, some of the Maf proteins (v-Maf, c-Maf and through their ability to regulate the expression and Nrl) can heterodimerize with c-Jun or c-Fos (Nishiza- function of cell cycle regulators such as Cyclin D1, p53, wa et al., 1989; Swaroop et al., 1992), whereas other p21cip1/waf1, p19ARF and p16.
    [Show full text]
  • Targeting the RB-E2F Pathway in Breast Cancer
    Oncogene (2016) 35, 4829–4835 © 2016 Macmillan Publishers Limited, part of Springer Nature. All rights reserved 0950-9232/16 www.nature.com/onc REVIEW Targeting the RB-E2F pathway in breast cancer J Johnson1, B Thijssen1, U McDermott2, M Garnett2, LFA Wessels1 and R Bernards1 Mutations of the retinoblastoma tumor-suppressor gene (RB1) or components regulating the CDK-RB-E2F pathway have been identified in nearly every human malignancy. Re-establishing cell cycle control through cyclin-dependent kinase (CDK) inhibition has therefore emerged as an attractive option in the development of targeted cancer therapy. The most successful example of this today is the use of the CDK4/6 inhibitor palbociclib combined with aromatase inhibitors for the treatment of estrogen receptor- positive breast cancers. Multiple studies have demonstrated that the CDK-RB-E2F pathway is critical for the control of cell proliferation. More recently, studies have highlighted additional roles of this pathway, especially E2F transcription factors themselves, in tumor progression, angiogenesis and metastasis. Specific E2Fs also have prognostic value in breast cancer, independent of clinical parameters. We discuss here recent advances in understanding of the RB-E2F pathway in breast cancer. We also discuss the application of genome-wide genetic screening efforts to gain insight into synthetic lethal interactions of CDK4/6 inhibitors in breast cancer for the development of more effective combination therapies. Oncogene (2016) 35, 4829–4835; doi:10.1038/onc.2016.32; published online 29 February 2016 INTRODUCTION multi-subunit protein complex containing partner (DP), RB-like, To maintain genome integrity, the mammalian cell cycle must be E2F and MuvB (DREAM) represses most if not all cell cycle gene 6 stringently regulated.
    [Show full text]
  • Aberrant Cytoplasmic Accumulation of Retinoblastoma Protein in Basal Cells May Lead to Increased Survival in Malignant Canine Mammary Tumours
    Original Paper Veterinarni Medicina, 59, 2014 (2): 76–80 Aberrant cytoplasmic accumulation of retinoblastoma protein in basal cells may lead to increased survival in malignant canine mammary tumours S. Gautam, N.K. Sood, K. Gupta Guru Angad Dev Veterinary and Animal Sciences University Ludhiana, Ludhiana, Punjab, India ABSTRACT: The retinoblastoma susceptibility geneRB-1 is a tumour suppressor gene that encodes a protein (Rb) that regulates the transition from the G1 phase to the S phase of the cell cycle. Inactivation of the Rb gene has been shown in a variety of human tumours, including breast, ovarian, hepatic, prostatic, and endometrial carcinomas. Although Rb protein is normally expressed in the nuclei of healthy cells, during carcinogenesis there is a partial or complete loss of nuclear expression. Recently, some reports have indicated aberrant cytoplasmic expression of Rb protein. However, little is known about its cytoplasmic expression and significance as a prognostic marker in canine mammary tumours (CMT). The present study was performed on 36 malignant CMT cases in order to assess the mutational status and prognostic significance of Rb in primary malignant CMT. We report an almost complete loss of nuclear expression of Rb protein with corresponding gain of aberrant cytoplasmic expression in basal/myoepithelial cells in CMT. Strikingly, our analysis reveals a significant positive correlation between survival time and cytoplasmic expression of Rb protein in basal cells. Moreover, cytoplasmic expression of Rb protein in basal cells was also correlated with tumour grade and stage. Keywords: canine mammary tumour; dogs; immunohistochemistry; retinoblastoma protein List of abbreviations CMT = canine mammary tumour, RB = retinoblastoma gene, Rb = retinoblastoma protein Breast cancer is the most frequent malignant tu- suppressors.
    [Show full text]
  • Dialogue Between Estrogen Receptor and E2F Signaling Pathways: the Transcriptional Coregulator RIP140 at the Crossroads*
    Advances in Bioscience and Biotechnology, 2013, 4, 45-54 ABB http://dx.doi.org/10.4236/abb.2013.410A3006 Published Online October 2013 (http://www.scirp.org/journal/abb/) Dialogue between estrogen receptor and E2F signaling pathways: The transcriptional coregulator RIP140 at the * crossroads Marion Lapierre1,2,3,4, Aurélie Docquier1,2,3,4, Audrey Castet-Nicolas1,2,3,4, Stéphan Jalaguier1,2,3,4, Catherine Teyssier1,2,3,4, Patrick Augereau1,2,3,4, Vincent Cavaillès1,2,3,4 1IRCM—Institut de Recherche en Cancérologie de Montpellier, Montpellier, France 2INSERM, U896, Montpellier, France 3Université Montpellier1, Montpellier, France 4Institut Régional du Cancer Montpellier, Montpellier, France Email: [email protected] Received 25 July 2013; revised 25 August 2013; accepted 19 September 2013 Copyright © 2013 Marion Lapierre et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. ABSTRACT Estrogen Receptors; Gene Expression; Cell Proliferation; Breast Cancer; Endocrine Therapies Estrogen receptors and E2F transcription factors are the key players of two nuclear signaling pathways which exert a major role in oncogenesis, particularly 1. ESTROGEN RECEPTOR AND E2F in the mammary gland. Different levels of dialogue SIGNALING PATHWAYS between these two pathways have been deciphered 1.1. Estrogen Signaling in Breast Cancer Cells and deregulation of the E2F pathway has been shown to impact the response of breast cancer cells to endo- Estrogens are steroid hormones that regulate growth and crine therapies. The present review focuses on the differentiation of a large number of target tissues such as transcriptional coregulator RIP140/NRIP1 which is the mammary gland, the reproductive tract and skeletal involved in several regulatory feed-back loops and and cardiovascular systems [1].
    [Show full text]
  • Androgens Repress Bcl-2 Expression Via Activation of the Retinoblastoma (RB) Protein in Prostate Cancer Cells
    Oncogene (2004) 23, 2161–2176 & 2004 Nature Publishing Group All rights reserved 0950-9232/04 $25.00 www.nature.com/onc Androgens repress Bcl-2 expression via activation of the retinoblastoma (RB) protein in prostate cancer cells Haojie Huang1, Ofelia L. Zegarra-Moro1, Douglas Benson1 and Donald J Tindall*,1 1Departments of Urology and Biochemistry/Molecular Biology, Mayo Clinic/Foundation, Rochester, MN 55905, USA The oncogene Bcl-2 is upregulated frequently in prostate function of the normal prostate gland, but also for the tumors following androgen ablation therapy,and Bcl-2 proliferation and survival of androgen-sensitive and - overexpression may contribute to the androgen-refractory refractory prostate cancer cells (Grossmann et al., 2001; relapse of the disease. However,the molecular mechanism Huang and Tindall, 2002; Zegarra-Moro et al., 2002). underlying androgenic regulation of Bcl-2 in prostate Surgical or pharmaceutical ablation of testicular andro- cancer cells is understood poorly. In this study,we gens has been the most effective treatment of metastatic demonstrated that no androgen response element (ARE) prostate cancer since 1941 (Huggins and Hodge, 1941). was identified in the androgen-regulated region of the P1 Both normal and malignant epithelial cells of the promoter of Bcl-2 gene,whereas,we provided evidence prostate undergo programmed cell death (apoptosis)in that the androgenic effect is mediated by E2F1 protein the absence of androgens (Isaacs, 1984; Kyprianou et al., through a putative E2F-binding site in the promoter. We 1990). However, a majority of prostate cancer patients further demonstrated that retinoblastoma (RB) protein usually relapse with tumors becoming refractory to plays a critical role in androgen regulation of Bcl-2.
    [Show full text]
  • Identification of a 60-Kilodalton Rb-Binding Protein, RBP60, That
    MOLECULARi AND CELLULAR BIOLOGY, OCt. 1992, p. 4327-4333 Vol. 12, No. 10 0270-7306/92/104327-07$02.00/0 Copyright © 1992, American Society for Microbiology Identification of a 60-Kilodalton Rb-Binding Protein, RBP60, That Allows the Rb-E2F Complex To Bind DNA SUBIR KUMAR RAY, MAY ARROYO, SRILATA BAGCHI, AND PRADIP RAYCHAUDHURI* Department ofBiochemistry (MIC 536), University ofIllinois at Chicago, Box 6998, Chicago, Illinois 60680 Received 11 March 1992/Returned for modification 17 April 1992/Accepted 1 July 1992 Several reports have indicated that the product of the retinoblastoma gene (Rb) complexes with the transcription factor E2F. We present evidence that the DNA-binding of the Rb-E2F complex involves another cellular factor. Addition of Rb to purified preparations of E2F does not generate an Rb-E2F complex that can bind DNA, and in fact, we see an inhibition of the DNA-binding ability of E2F. On the other hand, addition of Rb to cruder preparations of E2F results in the formation of an Rb-E2F complex (E2Fr) that can bind DNA and produces a distinct complex in gel retardation assays. We have identified and purified a 60-kDa protein that allows the Rb-E2F complex to bind DNA, and we show that this 60-kDa protein exerts its effect by directly interacting with Rb. The retinoblastoma gene is one of the best characterized contrast, Rb induces formation of a complex involving E2F tumor suppressor genes. The protein encoded by the reti- and its cognate sequence. By fractionating extracts from noblastoma gene, Rb, binds several DNA tumor virus onco- mouse L cells, we have identified and purified a factor, proteins (12, 15, 16) including adenovirus ElA, simian virus RBP60, that protects E2F from Rb inhibition and allows 40 T antigen, and papillomavirus E7 protein.
    [Show full text]
  • Rb and N-Ras Function Together to Control Differentiation in the Mouse
    Rb and N-ras Function Together to Control Differentiation in the Mouse The Harvard community has made this article openly available. Please share how this access benefits you. Your story matters Citation Takahashi, C., R. T. Bronson, M. Socolovsky, B. Contreras, K. Y. Lee, T. Jacks, M. Noda, R. Kucherlapati, and M. E. Ewen. 2003. “Rb and N-Ras Function Together To Control Differentiation in the Mouse.” Molecular and Cellular Biology 23 (15): 5256–68. doi:10.1128/ MCB.23.15.5256-5268.2003. Citable link http://nrs.harvard.edu/urn-3:HUL.InstRepos:41543053 Terms of Use This article was downloaded from Harvard University’s DASH repository, and is made available under the terms and conditions applicable to Other Posted Material, as set forth at http:// nrs.harvard.edu/urn-3:HUL.InstRepos:dash.current.terms-of- use#LAA MOLECULAR AND CELLULAR BIOLOGY, Aug. 2003, p. 5256–5268 Vol. 23, No. 15 0270-7306/03/$08.00ϩ0 DOI: 10.1128/MCB.23.15.5256–5268.2003 Copyright © 2003, American Society for Microbiology. All Rights Reserved. Rb and N-ras Function Together To Control Differentiation in the Mouse Chiaki Takahashi,1† Roderick T. Bronson,2,3 Merav Socolovsky,4 Bernardo Contreras,1 Kwang Youl Lee,1‡ Tyler Jacks,5 Makoto Noda,6 Raju Kucherlapati,7 and Mark E. Ewen1* Department of Medical Oncology and Medicine, Dana-Farber Cancer Institute and Harvard Medical School,1 and Rodent Histopathology Core3 and Harvard-Partners Center for Genetics and Genomics,7 Harvard Medical School, Boston, Massachusetts 02115; Department of Pathology, Tufts University Schools
    [Show full text]
  • Dissecting the DNA Binding Landscape and Gene Regulatory Network of P63 and P53
    bioRxiv preprint doi: https://doi.org/10.1101/2020.06.11.145540; this version posted June 12, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. Dissecting the DNA binding landscape and gene regulatory network of p63 and p53 Konstantin Riege1, Helene Kretzmer2, Simon S. McDade3, Steve Hoffmann1, Martin Fischer1,# 1 Computational Biology Group, Leibniz Institute on Aging – Fritz Lipmann Institute (FLI), Beutenbergstraße 11, 07745 Jena, Germany 2 Department of Genome Regulation, Max Planck Institute for Molecular Genetics, Ihnestraße 63-73, 14195 Berlin, Germany 3 Patrick G Johnston Centre for Cancer Research, Queen's University Belfast, 97 Lisburn Road, Belfast BT9 7AE, UK Running title: p63 GRN vs p53 GRN #To whom correspondence should be addressed. Email: [email protected] bioRxiv preprint doi: https://doi.org/10.1101/2020.06.11.145540; this version posted June 12, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. Abstract The transcription factor (TF) p53 is the best-known tumor suppressor, but its ancient sibling p63 (ΔNp63) is a master regulator of epidermis development and a key oncogenic driver in squamous cell carcinomas (SCC). Despite multiple gene expression studies becoming available in recent years, the limited overlap of reported p63-dependent genes has made it difficult to decipher the p63 gene regulatory network (GRN).
    [Show full text]