DOCSLIB.ORG

P21 Loss Cooperates with INK4 Inactivation Facilitating

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
P21 Loss Cooperates with INK4 Inactivation Facilitating Research Article p21 Loss Cooperates with INK4 Inactivation Facilitating Immortalization and Bcl-2–Mediated Anchorage- Independent Growth of Oncogene-Transduced Primary Mouse Fibroblasts Christopher J. Carbone,1 Xavier Gran˜a,1,2 E. Premkumar Reddy,1,2 and Dale S. Haines1,2 1Fels Institute for Cancer Research and Molecular Biology and 2Department of Biochemistry, Temple University School of Medicine, Philadelphia, Pennsylvania Abstract These kinase complexes phosphorylate pocket proteins pRb, p107, and p130, leading to release of the E2F family of transcriptional The INK4 and CIP cyclin-dependent kinase (Cdk) inhibitors regulators and transcription of genes required for cell cycle (CKI) activate pocket protein function by suppressing Cdk4 progression and DNA synthesis. Cdk activity is negatively regulated and Cdk2,respectively. Although these inhibitors are lost in by the INK4 (p16INK4A, p15INK4B, p18INK4C, and p19INK4D) and CIP/ tumors,deletion of individual CKIs results in modest Kip (p21Waf1/Cip1, p27Kip1, and p57Kip2) family of inhibitors (2). proliferation defects in murine models. We have evaluated Induction of these proteins by stress, growth-inhibitory, or cooperativity between loss of all INK4 family members (using differentiation-inducing signals provides a means of halting cell cdk4r24c mutant alleles that confer resistant to INK4 cycle progression to allow for repair or promoting exit from the cell inhibitors) and p21Waf1/Cip1 in senescence and transformation cycle. of mouse embryo fibroblasts (MEF). We show that mutant It was initially thought that Cdk4/Cdk6 and Cdk2 work cdk4r24c and p21 loss cooperate in pRb inactivation and MEF sequentially on pRb and play essential and nonredundant roles immortalization. Our studies suggest that cdk4r24c mediates in promoting cell cycle progression (3). However, the generation of resistance to p15INK4B/p16INK4A that accumulates over pas- knockout mouse models has challenged this notion and recent sage,whereas loss of p21 suppresses hyperoxia-induced Cdk2 results indicate that, at least for Cdk4 and Cdk2, they play inhibition and pRb dephosphorylation on MEF explantation in V12 overlapping roles in pRb phosphorylation and modulation of E2F- culture. Although cdk4r24c and p21 loss cooperate in H-ras / dependent gene expression in mouse fibroblasts (4). Cdk inhibitors c-myc–induced foci formation,they are insufficient for (CKI) from the INK4 and CIP/Kip families also possess overlapping oncogene-induced anchorage-independent growth. Interest- À/À V12 activities in regulating pocket protein function, although in ingly, p21 ; cdk4r24c MEFs expressing H-ras and c-myc contrast to Cdks, they promote pocket protein dephosphorylation display detachment-induced apoptosis and are transformed V12 and positively regulate their activities (5). by c-myc, H-ras ,and Bcl-2. We conclude that the INK4 Considering the fundamental roles of Cdks and CKIs in family and p21 loss cooperate in promoting pRb inactivation, V12 proliferation control, it is not surprising to find that the expression cell immortalization,and H-ras /c-myc–induced loss of and activity of these proteins are deregulated in cancer cells. For contact inhibition. In addition,absence of pRb function H-rasV12 c-myc example, human tumors frequently overexpress cyclin D1, Cdk4, renders + –transduced fibroblasts prone to and/or cyclin E. apoptosis when deprived of the extracellular matrix,and Conversely, expression of CKIs p15INK4B, p16INK4A, p18INK4C, and oncogene-induced anchorage-independent growth of pocket p27Kip1 are down-regulated in tumors (6). Consistent with the protein–deficient cells requires apoptotic suppression. [Cancer expression data, transgenic mice harboring elevated levels of cyclin Res 2007;67(9):4130–7] D1 or cyclin E are tumor prone, whereas those lacking individual CKIs are predisposed to cancer (7–9). For CKIs, however, the Introduction cancer predisposition phenotypes of single gene knockout are Quiescent cells require exposure to mitogenic growth factors to generally very mild and more dramatic phenotypes are observed in enter the cell cycle and pass through the restriction point. The key mice lacking multiple CKIs (6, 10–12). These studies reinforce the effectors promoting cell cycle entry and progression through the idea of functional overlap between CKI proteins and suggest that G1-Stransition are the cyclin-dependent kinase (Cdk) 4/Cdk6- inactivation or down-regulation of multiple CKIs will be required cyclin D and Cdk2-cyclin E kinase complexes (1). Cdk4/Cdk6 for cellular transformation and tumor development. associates with mitogen-induced D-type cyclins in early G1, Because of the difficulties in studying the cellular transformation whereas Cdk2 pairs with cyclin Es that accumulate in late G1. process in vivo, cell-based models using primary mouse embryo fibroblasts (MEF) have been extensively used to define genes regulating ‘‘normal’’ cell proliferation and how gene disruption affects various growth characteristics in culture. Although there are Note: Supplementary data for this article are available at Cancer Research Online clear differences between the proliferative characteristics of human (http://cancerres.aacrjournals.org/). and murine fibroblasts, studies have defined genes that are Requests for reprints: Dale S. Haines, Fels Institute for Cancer Research and Molecular Biology and Department of Biochemistry, Temple University School of required for the escape of culture-induced growth arrest Medicine, Philadelphia, PA 19140. Phone: 215-707-5765; Fax: 215-707-2102; E-mail: (or senescence), oncogene-induced proliferation, and anchorage- [email protected]. À/À I2007 American Association for Cancer Research. independent growth (13). For example, p53 MEFs do not V12 doi:10.1158/0008-5472.CAN-07-0499 undergo culture or oncogenic H-ras –induced proliferation arrest Cancer Res 2007; 67: (9). May 1, 2007 4130 www.aacrjournals.org Downloaded from cancerres.aacrjournals.org on September 30, 2021. © 2007 American Association for Cancer Research. cdk4r24c and p21 Loss Cooperate in MEF Transformation and are susceptible to H-rasV12–mediated anchorage-independent activity was determined from immunopurified complexes as described growth (14–16). Interestingly, deletion of pRb and p107, or all three previously (24). B h pocket proteins, allows MEFs to proliferate on introduction of Senescence-associated -galactosidase assay. -galactosidase activity mutant RASalone yet confers inefficient (at least when compared was detected as described previously (25) with slight modifications. MEFs À À from passage 10 of the 3T3 protocol were washed once with PBS, fixed with with p53 / MEFs) anchorage-independent proliferation after 0.5% glutaraldehyde (PBS), and washed in PBS supplemented with 1 mmol/L transduction with this and other oncogenes, including c-myc MgCl2. MEFs were stained with X-gal solution [1 mg/mL X-gal, 0.12 mmol/L (17). In terms of CKIs, MEFs lacking individual INKs or CIPs/Kips K3Fe(CN)6, 0.12 mmol/L Fe4(CN)6, and 1 mmol/L MgCl2 in PBS(pH 6.0)] at display only modest, if any, predisposition to immortalization in 37jC. V12 V12 culture, H-ras –induced proliferation, or H-ras + c-myc– Oncogene-induced foci and soft agar assays. Phoenix cells were mediated anchorage-independent growth (6, 10). Thus, a common transfected with 10 Ag of each retroviral plasmid using the CalPhos theme that emerges from these studies is that in murine Mammalian Transfection kit (BD Biosciences) and 25 Amol/L chloroquine fibroblasts, the CKIs are likely to play overlapping roles in (Sigma). The medium was changed 24 h after the transfection. Forty-eight suppressing culture and oncogene-induced proliferative arrest hours after transfection, the retrovirus-containing medium was collected, A and cells lacking pRb function will require inactivation or filtered, supplemented with 5 g/mL polybrene (Sigma), and used to infect early-passage MEFs growing in 3% oxygen/5% CO (done in duplicate for activation of other pathways to allow for oncogene-induced 2 each genotype). Infected cell populations were selected with 2 Ag/mL anchorage-independent proliferation. puromycin. For foci formation assays, 3 Â 105 puromycin-selected cells were Mutation of residue 24 from an arginine to cysteine in Cdk4 seeded into 10-cm plates. Cells were maintained for 10 days in a standard occurs in familial cases of melanomas and confers resistance of the tissue culture environment and the medium was changed every 2 days. For protein to inhibition by INK4 family members (18, 19). Mouse soft agar assays, MEFs were sequentially transduced with the indicated modeling experiments have shown that mice harboring a knock-in viruses, and puromycin-selected and 4 Â 104 cells were placed into 0.38% cdk4r24c mutation develop tumors at an accelerated rate (18–20). Nobel agar in DMEM containing 10% fetal bovine serum. This was overlaid Moreover, cdk4r24c MEFs are resistant to culture-induced senes- on solidified 0.7% Nobel agar containing culture medium in a 6-cm dish. cence and are prone to transformation by oncogenes. However, The cells were fed weekly and colonies were evaluated after 2 weeks. Viable these phenotypes are again mild, especially when compared with colonies were stained with 0.05% nitroblue tetrazolium. Detachment-induced apoptosis studies. The detachment-induced MEFs lacking all pocket proteins (21, 22), raising the possibility of apoptosis assays were done as described previously (26). Briefly, equal functional redundancy between INK4 proteins and
Load more

Recommended publications
  • INK4 Locus of the Tumor-Resistant Rodent, the Naked Mole Rat, Expresses a Functional P15/P16 Hybrid Isoform
    INK4 locus of the tumor-resistant rodent, the naked mole rat, expresses a functional p15/p16 hybrid isoform Xiao Tiana,1, Jorge Azpuruaa,1,2, Zhonghe Kea, Adeline Augereaub, Zhengdong D. Zhangc, Jan Vijgc, Vadim N. Gladyshevb, Vera Gorbunovaa,3, and Andrei Seluanova,3 aDepartment of Biology, University of Rochester, Rochester, NY 14627; bBrigham and Women’s Hospital, Harvard Medical School, Boston, MA 02115; and cAlbert Einstein College of Medicine, Bronx, NY 10461 Edited* by Eviatar Nevo, Institute of Evolution, Haifa, Israel, and approved December 1, 2014 (received for review September 21, 2014) The naked mole rat (Heterocephalus glaber) is a long-lived and because, in mammals, it encodes three distinct tumor suppressors: tumor-resistant rodent. Tumor resistance in the naked mole rat p16INK4a, p15INK4b, and p19ARF (p14ARF in human) (9). These is mediated by the extracellular matrix component hyaluronan three proteins coordinate a signaling network that depends on of very high molecular weight (HMW-HA). HMW-HA triggers hy- the activities of the retinoblastoma protein (RB) and the p53 persensitivity of naked mole rat cells to contact inhibition, which is tumor suppressor protein. The p16INK4a and p15INK4b pro- associated with induction of the INK4 (inhibitors of cyclin depen- teins are cyclin-dependent kinase inhibitors that directly in- dent kinase 4) locus leading to cell-cycle arrest. The INK4a/b locus is hibit the binding of cyclins to their target cyclin-dependent among the most frequently mutated in human cancer. This locus kinases (10). p16INK4a is involved in establishing replicative encodes three distinct tumor suppressors: p15INK4b, p16INK4a, and senescence, oncogene-induced senescence, and stress-induced INK4b ARF (alternate reading frame).
    [Show full text]
  • The Involvement of Ubiquitination Machinery in Cell Cycle Regulation and Cancer Progression
    International Journal of Molecular Sciences Review The Involvement of Ubiquitination Machinery in Cell Cycle Regulation and Cancer Progression Tingting Zou and Zhenghong Lin * School of Life Sciences, Chongqing University, Chongqing 401331, China; [email protected] * Correspondence: [email protected] Abstract: The cell cycle is a collection of events by which cellular components such as genetic materials and cytoplasmic components are accurately divided into two daughter cells. The cell cycle transition is primarily driven by the activation of cyclin-dependent kinases (CDKs), which activities are regulated by the ubiquitin-mediated proteolysis of key regulators such as cyclins, CDK inhibitors (CKIs), other kinases and phosphatases. Thus, the ubiquitin-proteasome system (UPS) plays a pivotal role in the regulation of the cell cycle progression via recognition, interaction, and ubiquitination or deubiquitination of key proteins. The illegitimate degradation of tumor suppressor or abnormally high accumulation of oncoproteins often results in deregulation of cell proliferation, genomic instability, and cancer occurrence. In this review, we demonstrate the diversity and complexity of the regulation of UPS machinery of the cell cycle. A profound understanding of the ubiquitination machinery will provide new insights into the regulation of the cell cycle transition, cancer treatment, and the development of anti-cancer drugs. Keywords: cell cycle regulation; CDKs; cyclins; CKIs; UPS; E3 ubiquitin ligases; Deubiquitinases (DUBs) Citation: Zou, T.; Lin, Z. The Involvement of Ubiquitination Machinery in Cell Cycle Regulation and Cancer Progression. 1. Introduction Int. J. Mol. Sci. 2021, 22, 5754. https://doi.org/10.3390/ijms22115754 The cell cycle is a ubiquitous, complex, and highly regulated process that is involved in the sequential events during which a cell duplicates its genetic materials, grows, and di- Academic Editors: Kwang-Hyun Bae vides into two daughter cells.
    [Show full text]
  • Increased Expression of Unmethylated CDKN2D by 5-Aza-2'-Deoxycytidine in Human Lung Cancer Cells
    Oncogene (2001) 20, 7787 ± 7796 ã 2001 Nature Publishing Group All rights reserved 0950 ± 9232/01 $15.00 www.nature.com/onc Increased expression of unmethylated CDKN2D by 5-aza-2'-deoxycytidine in human lung cancer cells Wei-Guo Zhu1, Zunyan Dai2,3, Haiming Ding1, Kanur Srinivasan1, Julia Hall3, Wenrui Duan1, Miguel A Villalona-Calero1, Christoph Plass3 and Gregory A Otterson*,1 1Division of Hematology/Oncology, Department of Internal Medicine, The Ohio State University-Comprehensive Cancer Center, Columbus, Ohio, OH 43210, USA; 2Department of Pathology, The Ohio State University-Comprehensive Cancer Center, Columbus, Ohio, OH 43210, USA; 3Division of Human Cancer Genetics, Department of Molecular Virology, Immunology and Medical Genetics, The Ohio State University-Comprehensive Cancer Center, Columbus, Ohio, OH 43210, USA DNA hypermethylation of CpG islands in the promoter Introduction region of genes is associated with transcriptional silencing. Treatment with hypo-methylating agents can Methylation of cytosine residues in CpG sequences is a lead to expression of these silenced genes. However, DNA modi®cation that plays a role in normal whether inhibition of DNA methylation in¯uences the mammalian development (Costello and Plass, 2001; expression of unmethylated genes has not been exten- Li et al., 1992), imprinting (Li et al., 1993) and X sively studied. We analysed the methylation status of chromosome inactivation (Pfeifer et al., 1990). To date, CDKN2A and CDKN2D in human lung cancer cell lines four mammalian DNA methyltransferases (DNMT) and demonstrated that the CDKN2A CpG island is have been identi®ed (Bird and Wole, 1999). Disrup- methylated, whereas CDKN2D is unmethylated. Treat- tion of the balance in methylated DNA is a common ment of cells with 5-aza-2'-deoxycytidine (5-Aza-CdR), alteration in cancer (Costello et al., 2000; Costello and an inhibitor of DNA methyltransferase 1, induced a dose Plass, 2001; Issa et al., 1993; Robertson et al., 1999).
    [Show full text]
  • Structure of Human Cyclin-Dependent Kinase Inhibitor P19ink4d
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector Research Article 1279 Structure of human cyclin-dependent kinase inhibitor p19INK4d: comparison to known ankyrin-repeat-containing structures and implications for the dysfunction of tumor suppressor p16INK4a Roland Baumgartner1, Carlos Fernandez-Catalan1, Astar Winoto2, Robert Huber1, Richard A Engh1* and Tad A Holak1* Background: The four members of the INK4 gene family (p16INK4a, p15INK4b, Addresses: 1Max Planck Institute for Biochemistry, p18INK4c and p19INK4d) inhibit the closely related cyclin-dependent kinases D-82152 Martinsried, Federal Republic of Germany 2 CDK4 and CDK6 as part of the regulation of the G →S transition in the cell- and Department of Molecular and Cell Biology, 1 University of California Berkeley, CA 94720-3200, INK4a division cycle. Loss of INK4 gene product function, particularly that of p16 , USA. is found in 10–60% of human tumors, suggesting that broadly applicable anticancer therapies might be based on restoration of p16INK4a CDK inhibitory *Corresponding authors. function. Although much less frequent, defects of p19INK4d have also been E-mail: [email protected] [email protected] associated with human cancer (osteosarcomas). The protein structures of some INK4 family members, determined by nuclear magnetic resonance (NMR) Key words: cell cycle, CDK4 inhibitor, p19INK4d, spectroscopy and X-ray techniques, have begun to clarify the functional role of p16INK4a, structure p16INK4a and the dysfunction introduced by the mutations associated with Received: 9 April 1998 human tumors. Revisions requested: 5 June 1998 Revisions received: 30 July 1998 Results: The crystal structure of human p19INK4d has been determined at 1.8 Å Accepted: 31 July 1998 resolution using multiple isomorphous replacement methods.
    [Show full text]
  • Induction of Cip/Kip and Ink4 Cyclin Dependent Kinase Inhibitors by Interferon-A in Hematopoietic Cell Lines
    Oncogene (1997) 14, 415 ± 423 1997 Stockton Press All rights reserved 0950 ± 9232/97 $12.00 Induction of Cip/Kip and Ink4 cyclin dependent kinase inhibitors by interferon-a in hematopoietic cell lines Olle Sangfelt, Sven Erickson, Stefan Einhorn and Dan Grande r Department of Oncology/Pathology, Radiumhemmet, Building P8, Karolinska Hospital and Institute, S-171 76 Stockholm, Sweden One prominent eect of IFNs is their cell growth exert cell growth inhibitory eects. It has since been inhibitory activity. The exact molecular mechanism found that this antiproliferative action of IFNs can be behind this inhibition of proliferation remains to be observed in many cell types from established cell lines, elucidated. Possible eectors for IFN-induced growth normal tissues and primary tumor cells (Taylor- inhibition are the recently discovered cyclin-dependent Papadimitriou and Rozengurt, 1985). It has been kinase inhibitors. The eect of IFN-a treatment on the postulated that this eect may, at least in part, members of the Ink4 and Cip/Kip families of Cdk mediate the antitumor eects observed in some inhibitors was investigated in three hematopoietic cell malignancies (Brenning et al., 1985). In vitro culture lines Daudi, U-266 and H9. Two of these cell lines, of susceptible cell types with IFN commonly leads to Daudi and U-266, respond to IFN-a by G1 arrest, G1 arrest, but sometimes to a lengthening of S-phase or whereas the H9 cell line is not growth arrested by IFN-a. all phases of the cell cycle (Roos et al., 1984). We show that a p53-independent upregulation of p21 With the discovery of the proteins that normally mRNA occurs following IFN-a treatment in all three cell regulate the cell cycle, some of the molecular eectors lines.
    [Show full text]
  • The Role of Cdks and Cdkis in Murine Development
    International Journal of Molecular Sciences Review The Role of CDKs and CDKIs in Murine Development Grace Jean Campbell , Emma Langdale Hands and Mathew Van de Pette * Epigenetic Mechanisms of Toxicology Lab, MRC Toxicology Unit, Cambridge University, Cambridge CB2 1QR, UK; [email protected] (G.J.C.); [email protected] (E.L.H.) * Correspondence: [email protected] Received: 8 July 2020; Accepted: 26 July 2020; Published: 28 July 2020 Abstract: Cyclin-dependent kinases (CDKs) and their inhibitors (CDKIs) play pivotal roles in the regulation of the cell cycle. As a result of these functions, it may be extrapolated that they are essential for appropriate embryonic development. The twenty known mouse CDKs and eight CDKIs have been studied to varying degrees in the developing mouse, but only a handful of CDKs and a single CDKI have been shown to be absolutely required for murine embryonic development. What has become apparent, as more studies have shone light on these family members, is that in addition to their primary functional role in regulating the cell cycle, many of these genes are also controlling specific cell fates by directing differentiation in various tissues. Here we review the extensive mouse models that have been generated to study the functions of CDKs and CDKIs, and discuss their varying roles in murine embryonic development, with a particular focus on the brain, pancreas and fertility. Keywords: cyclin-dependent kinase; CDK inhibitors; mouse; development; knock-out models 1. Introduction Cyclin-dependent kinases (CDKs) are proteins that, by definition, require the binding of partner cyclin proteins in order to phosphorylate a series of target proteins.
    [Show full text]
  • Targeting Dysregulation of the Cyclin D-Cdk4/6-Ink4-Rb Pathway ©2017 Anis 338
    Journal of Cancer Prevention & Current Research Editorial Open Access Targeting dysregulation of the cyclin d-cdk4/6-ink4- rb pathway Introduction Volume 8 Issue 5 - 2017 The cyclin D-cyclin dependent kinase (CDK) 4/6-inhibitor of Hajj Adel Anis CDK4 (INK4)-retinoblastoma (Rb) pathway controls cell cycle Cedars - Jebel Ali International Hospital, UAE progression by regulating the G1-S checkpoint. Dysregulation of the cyclin D-CDK4/6-INK4-Rb pathway results in increased Correspondence: Hajj Adel Anis, Medical Oncologist, Cedars- Jebel Ali International Hospital, 9370 Rue Lajeunesse, Montreal, 1,2 proliferation, and is frequently observed in many types of cancer. UAE, Tel 438 992 5516, Email Thus, the development of selective CDK4/6 inhibitors offers a novel therapeutic approach for patients with advanced cancer. Received: August 07, 2017 | Published: September 28, 2017 The rationale for inhibiting CDK4/6 To appreciate selective targeting in cancer, it is important to understand that the cell cycle exists in 4 phases: G0/G1, synthesis (S), G2, and mitosis (M). In G0, the cell is in an arrested state that can be either permanent (after senescence or terminal differentiation) or quiescent until mitogenic factors stimulate the cell to reenter the cell cycle. In the G1phase, these mitogenic factors activate intracellular signaling events that drive the cell cycle to progress into S phase. In the S phase, DNA is replicated. In the G2 phase, protein synthesis and cell growth occur, and in the M phase, the cell divides into 2 daughter cells. Progression through these 4 phases is regulated by members of This multifactorial process presents research motivation for drug the CDK family, which includes CDKs 1, 2, 4, and 6.
    [Show full text]
  • Novel INK4 Proteins, P19 and P18, Are Specific Inhibitors of the Cyclin
    MOLECULAR AND CELLULAR BIOLOGY, May 1995, p. 2672–2681 Vol. 15, No. 5 0270-7306/95/$04.0010 Copyright q 1995, American Society for Microbiology Novel INK4 Proteins, p19 and p18, Are Specific Inhibitors of the Cyclin D-Dependent Kinases CDK4 and CDK6 HIROSHI HIRAI,1 MARTINE F. ROUSSEL,1 JUN-YA KATO,1 RICHARD A. ASHMUN,1,2 1,3 AND CHARLES J. SHERR * Departments of Tumor Cell Biology1 and Experimental Oncology2 and Howard Hughes Medical Institute,3 St. Jude Children’s Research Hospital, Memphis, Tennessee 38105 Received 13 January 1995/Returned for modification 14 February 1995/Accepted 22 February 1995 Cyclin D-dependent kinases act as mitogen-responsive, rate-limiting controllers of G1 phase progression in mammalian cells. Two novel members of the mouse INK4 gene family, p19 and p18, that specifically inhibit the kinase activities of CDK4 and CDK6, but do not affect those of cyclin E-CDK2, cyclin A-CDK2, or cyclin B-CDC2, were isolated. Like the previously described human INK4 polypeptides, p16INK4a/MTS1 and p15INK4b/MTS2, mouse p19 and p18 are primarily composed of tandemly repeated ankyrin motifs, each ca. 32 amino acids in length. p19 and p18 bind directly to CDK4 and CDK6, whether untethered or in complexes with D cyclins, and can inhibit the activity of cyclin D-bound cyclin-dependent kinases (CDKs). Although neither protein interacts with D cyclins or displaces them from preassembled cyclin D-CDK complexes in vitro, both form complexes with CDKs at the expense of cyclins in vivo, suggesting that they may also interfere with cyclin-CDK assembly.
    [Show full text]
  • Human CDKN2D / P19ink4d Protein (GST Tag)
    Human CDKN2D / p19ink4d Protein (GST Tag) Catalog Number: 12558-H09E General Information SDS-PAGE: Gene Name Synonym: INK4D; p19; p19-INK4D Protein Construction: A DNA sequence encoding the human CDKN2D (P55273) (Met 10Leu 166) was fused with the GST tag at the N-terminus. Source: Human Expression Host: E. coli QC Testing Purity: > 90 % as determined by SDS-PAGE Bio Activity： Protein Description Immobilized human GST-CDKN2D at 10 μg/ml (100 μl/well) can bind biotinylated human GST-CDK4 (Cat:10732-H09B), The EC50 of Cyclin-dependent kinase inhibitor 2D(also known as CDKN2D or p19ink4d), biotinylated human GST-CDK4 (Cat:10732-H09B) is 0.52-1.2 μg/ml. a member of the INK4 family of cyclin-dependent kinase (CDK) inhibitors, negatively regulates the cyclin D-CDK4/6 complexes, which promote G1/S Endotoxin: transition by phosphorylating the retinoblastoma tumor-suppressor gene product. It is clearly shown that DNA repair is the main target of p19ink4d Please contact us for more information. effect and that diminished apoptosis is a downstream event. Experiments has uncovered a role of p19INK4d as a regulator of DNA-damage-induced Stability: apoptosis and suggest that it protects cells from undergoing apoptosis by Samples are stable for up to twelve months from date of receipt at -70 ℃ allowing a more efficient DNA repair. It has been demonstrated that p19INK4d expression enhances cell survival under genotoxic conditions. Predicted N terminal: Met Previous work has shown that inactivation of the cyclin-dependent kinase inhibitor (CKI) p19(Ink4d) leads to progressive hearing loss attributable to Molecular Mass: inappropriate DNA replication and subsequent apoptosis of hair cells.
    [Show full text]
  • The Role of Polo-Like Kinase 1 in Carcinogenesis: Cause Or Consequence?
    Published OnlineFirst November 21, 2013; DOI: 10.1158/0008-5472.CAN-13-2197 Cancer Review Research The Role of Polo-like Kinase 1 in Carcinogenesis: Cause or Consequence? Brian D. Cholewa1,2, Xiaoqi Liu4, and Nihal Ahmad1,2,3 Abstract Polo-like kinase 1 (Plk1) is a well-established mitotic regulator with a diverse range of biologic functions continually being identified throughout the cell cycle. Preclinical evidence suggests that the molecular targeting of Plk1 could be an effective therapeutic strategy in a wide range of cancers; however, that success has yet to be translated to the clinical level. The lack of clinical success has raised the question of whether there is a true oncogenic addiction to Plk1 or if its overexpression in tumors is solely an artifact of increased cellular proliferation. In this review, we address the role of Plk1 in carcinogenesis by discussing the cell cycle and DNA damage response with respect to their associations with classic oncogenic and tumor suppressor pathways that contribute to the transcriptional regulation of Plk1. A thorough examination of the available literature suggests that Plk1 activity can be dysregulated through key transformative pathways, including both p53 and pRb. On the basis of the available literature, it may be somewhat premature to draw a definitive conclusion on the role of Plk1 in carcinogenesis. However, evidence supports the notion that oncogene dependence on Plk1 is not a late occurrence in carcinogenesis and it is likely that Plk1 plays an active role in carcinogenic transformation. Cancer Res; 73(23); 6848–55. Ó2013 AACR. Introduction of directly contributing to carcinogenesis.
    [Show full text]
  • P16ink4a Overexpression in Cancer: a Tumor Suppressor Gene Associated with Senescence and High-Grade Tumors
    Oncogene (2011) 30, 2087–2097 & 2011 Macmillan Publishers Limited All rights reserved 0950-9232/11 www.nature.com/onc REVIEW p16Ink4a overexpression in cancer: a tumor suppressor gene associated with senescence and high-grade tumors C Romagosa1, S Simonetti2,LLo´pez-Vicente1, A Mazo3, ME Lleonart1, J Castellvi1 and S Ramon y Cajal1 1Pathology Department, Fundacio´ Institut de Recerca, Hospital Universitari Vall d’Hebron, Universitat Auto`noma de Barcelona, Barcelona, Spain; 2Pangaea Biotech, Oncology Laboratory, USP Dexeus University Institute, Barcelona, Spain and 3Department of Biochemistry and Molecular Biology, University of Barcelona, Institute of Biomedicine (IBUB), Barcelona, Spain p16Ink4a is a protein involved in regulation of the cell cycle. Physiological role of p16Ink4a Currently, p16Ink4a is considered a tumor suppressor protein because of its physiological role and downregulated expres- p16Ink4a and the cell cycle sion in a large number of tumors. Intriguingly, overexpression p16Ink4a is the principal member of the Ink4 family of p16Ink4a has also been described in several tumors. This of CDK inhibitors. It is codified by a gene localized review attempts to elucidate when and why p16Ink4a over- on chromosome 9p21 within the INK4a/ARF locus, expression occurs, and to suggest possible implications of which encodes for two different proteins with different p16Ink4a in the diagnosis, prognosis and treatment of cancer. promoters: p16Ink4a and p19ARF. Both proteins have Oncogene (2011) 30, 2087–2097; doi:10.1038/onc.2010.614; antiproliferative biological activity, and are involved in published online 7 February 2011 the retinoblastoma protein (Rb) and p53 pathways, respec- tively (Serrano, 1997; Weber et al., 2000; Pei and Xiong, Keywords: p16Ink4a; overexpression; cancer; senescence; 2005).
    [Show full text]
  • Expression of P27kip1, P21waf1 and P53 Does Not Correlate with Prognosis in Node-Negative Invasive Ductal Carcinoma of the Breast
    ANTICANCER RESEARCH 26: 1657-1668 (2006) Expression of p27KIP1, p21WAF1 and p53 does Not Correlate with Prognosis in Node-negative Invasive Ductal Carcinoma of the Breast HELEN P. KOUREA1, ANGELOS K. KOUTRAS2, VASSILIKI ZOLOTA1, IRENE GRIMANI3, EVANGELOS TZORACOELEFTHERAKIS4, DIMITRIOS KOUKOURAS4, GEORGE FOUNTZILAS5, HELEN GOGAS6, KYRIAKOS KYRIAKOU7, ADAMOS ADAMOU7 and HARALAMBOS P. KALOFONOS2 Departments of 1Pathology, 2Internal Medicine and 4Surgery, University Hospital of Patras, Patras; 3Department of Biostatistics, Hellenic Cooperative Oncology Group Data Office, Athens; 5Aristotle University of Thessaloniki School of Medicine, Thessaloniki; 6First Department of Medicine, University of Athens, Athens, Greece; 7The Cyprus Institute of Neurology and Genetics, Nicosia, Cyprus Abstract. The expressions of p27Kip1 (p27) and p21Waf1 (p21) generally has a favorable outcome even without adjuvant cyclin-dependent kinase inhibitors and p53 were examined in a therapy, 30% of the patients will die of their disease without series of 170 node-negative breast carcinomas (NNBCs) to adjuvant treatment (7). Since the classic clinicopathological evaluate their prognostic significance. Low nuclear (p27TN) parameters are often not adequate to identify these patients, and cytoplasmic (p27TC) p27 expressions were noted in 66% and the benefit of adjuvant therapy in this setting is debatable and 81% of NNBCs, respectively. p21 and p53 overexpressions (7), additional information provided by novel prognostic were detected in 56% and 26%, respectively. Low p27TN was factors will assist in the fine-tuning of the prognostication significantly associated with high grade (p=0.001), age ≤50 process and the treatment decision-making. years (p=0.01), negative hormone receptors (p<0.001), low Factors involved in cell cycle control have been shown to be p27TC (p<0.001) and p53 overexpression (p=0.02).
    [Show full text]