DOCSLIB.ORG

Regulation of the Transcription Factor YY1 by Phosphorylation Ari Kassardjian

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Regulation of the Transcription Factor YY1 by Phosphorylation Ari Kassardjian Florida State University Libraries Electronic Theses, Treatises and Dissertations The Graduate School 2012 Regulation of the Transcription Factor YY1 by Phosphorylation Ari Kassardjian Follow this and additional works at the FSU Digital Library. For more information, please contact [email protected] FLORIDA STATE UNIVERSITY COLLEGE OF ARTS AND SCIENCES REGULATION OF THE TRANSCRIPTION FACTOR YY1 BY PHOSPHORYLATION By ARI KASSARDJIAN A Dissertation submitted to the Program of Molecular Biophysics in partial fulfillment of the requirements for the degree of Doctor of Philosophy Degree Awarded: Summer Semester, 2012 Ari Kassardjian defended this dissertation on June 21, 2012. The members of the supervisory committee were: Myra M. Hurt Professor Directing Dissertation Akash Gunjan University Representative Lloyd Epstein Committee Member Cathy Levenson Committee Member Hank Bass Committee Member The Graduate School has verified and approved the above-named committee members, and certifies that the dissertation has been approved in accordance with university requirements. ii ACKNOWLEDGMENTS I feel very lucky and privileged that I had the opportunity to work in the laboratory of Dr. Myra Hurt; I am forever grateful for the support and guidance that I have received from my advisor. I am honored to have worked under the supervision of a great mentor and I cannot thank her enough! I would like to thank the members of my committee: Dr. Akash Gunjan, Dr. Lloyd Epstein, Dr. Hank Bass, and Dr. Cathy Levenson. Without their valuable feedback and accessibility, this work would not have been achieved. This work would have not been possible without Beth Alexander, our lab director. She taught me many techniques and was always there to help me with all my work. I would also like to thank Dr. Raed Rizkallah, an amazing researcher who led by example. He was always present and available to help answer my questions and guide me in the right direction. I like to thank Dr. Sarah Riman for being a great friend and colleague in the lab. Beth, Raed and Sarah had major, direct and indirect, contributions to this study and I will never forget our experiences together. Also, I would like to thank all the faculty members, staff, colleagues and friends in the Department of Biomedical Sciences and the Molecular Biophysics Graduate program at Florida State University. Lastly, I would like to thank my parents, Chahe and Rosine, as well as my brother, Araz for their unwavering love, unending support and encouragement. iii TABLE OF CONTENTS LIST OF TABLES………………………………………………………………………..v LIST OF FIGURES……………………………………………………………………...vi ABSTRACT…………………………………………………………………………….viii CHAPTER 1 INTRODUCTION ........................................................................... 1 DISCOVERY AND CHARACTERIZATION OF YY1 ......................................... 2 YY1 AND CELL CYCLE REGULATION ........................................................... 7 YY1 REGULATION ........................................................................................... 9 POSTTRANSLATIONAL REGULATION OF YY1 ........................................... 11 KINASES THAT PHOSPHORYLATE YY1 ...................................................... 24 GOALS OF THE PROJECT ............................................................................ 28 CHAPTER 2 AURORA B KINASE PHOSPHORYLATES THE TRANSCRIPTION FACTOR YY1 AT G2/M TRANSITION OF THE CELL CYCLE AND MODULATES ITS TRANSCRIPTIONAL ACTIVITY ................................................................. 29 INTRODUCTION .................................................................................................. 30 MATERIALS AND METHODS ................................................................................. 31 RESULTS .......................................................................................................... 39 DISCUSSION ..................................................................................................... 59 CHAPTER 3 IDENTIFICATION OF TYROSINE KINASES THAT PHOSPHORYLATE YY1 IN VITRO ................................................................................................... 65 INTRODUCTION .................................................................................................. 65 MATERIAL AND METHODS .................................................................................. 66 RESULTS .......................................................................................................... 72 DISCUSSION ..................................................................................................... 87 CHAPTER 4 CONCLUSIONS .......................................................................... 92 APPENDIX ......................................................................................................... 95 REFERENCES ................................................................................................... 97 BIOGRAPHICAL SKETCH .............................................................................. 114 iv LIST OF TABLES Table I. Known in vivo Posttranslational modifications of YY1. ..................... 16 Table II. In vivo phosphorylation sites of YY1 ......................................... ……22 v LIST OF FIGURES Figure 1. Diagram of the different functional domains of human YY1 and the location of each phosphorylated amino acid residue. ................................................ 4 Figure 2. Structure of the YY1 cocrystal structure. ...................................................... 7 Figure 3. Protein phosphorylation and protein dephosphorylation regulate the activities of transcription factors through multiple mechanisms. ................. 17 Figure 4. Phosphorylation of YY1 in nocodazole blocked extracts is detected by antiphospho-S184 antibody.. ...................................................................... 42 Figure 5. Serine 184 phosphorylation on YY1 peaks at the G2/M stage of the cell cycle. .......................................................................................................... 45 Figure 6. Aurora B phosphorylates YY1 at serine 184 in vitro. .................................. 47 Figure 7. PKA and ROCK1 phosphorylate YY1 at serine 184 in vitro. ....................... 49 Figure 8. Aurora B phosphorylates YY1 at serine 184 in vivo. ................................... 51 Figure 9. Cell cycle analysis and cellular localization of YY1 phospho-mutants in HEK293 cells.. ............................................................................................ 53 Figure 10. Phosphorylation of YY1 in its regulatory domain modulates transcriptional activity......................................................................................................... 55 Figure 11. YY1 S180,184D phospho-mutant exhibits an increased DNA binding affinity in vitro.. ............................................................................................ 56 Figure 12. The histone acetyltransferase p300 acetylates full length YY1 wild type in vitro. ............................................................................................................ 58 Figure 13. Schematic model of the regulation of YY1 by Aurora B at G2/M.. ............... 64 Figure 14. YY1 is phosphorylated on tyrosine residue(s) in vitro and in vivo.. ............. 73 Figure 15. YY1 is phosphorylated on tyrosine residues during mitosis.. ...................... 75 Figure 16. Structural and functional domains of YY1 showing the location of the six tyrosine residues in YY1.. ........................................................................... 77 Figure 17. Phosphorylation of Y251 and Y254 by mitotic extracts. .............................. 79 Figure 18. Tyrosine kinases included in the kinase profiling screen. ............................. 81 vi Figure 19. Multiple tyrosine kinases phosphorylate YY1 in vitro .................................. 82 Figure 20. Dot blot assay of non-phosphorylated and tyrosine phosphorylated synthetic peptides of YY1 probed with anti-pY251 or anti-pY254. ............................. 83 Figure 21. FAK and SRC phosphorylate YY1 at Y251 and Y254 in vitro ..................... 84 Figure 22. Protein expression and DNA binding activity of EGFP-YY1 Y251F and Y254F in HeLa extracts. ............................................................................. 85 Figure 23. Examples of zinc finger containing proteins with conserved tyrosine (Y) or phenylalanine (F) residues at the linker region ........................................... 86 Figure 24. Protein expression and DNA binding activity of EGFP-YY1 Y383F in HeLa extracts.. ..................................................................................................... 87 vii ABSTRACT Reversible protein phosphorylation plays an integral role in the regulation of eukaryotic cellular signaling, especially for transcription factors. Yin Yang 1 (YY1) is a ubiquitously expressed and highly conserved multifunctional transcription factor that is involved in a variety of cellular processes. Many YY1-regulated genes have crucial roles in cell proliferation, differentiation, apoptosis, and cell cycle regulation. Numerous mechanisms have been shown to regulate the function of YY1, such as DNA binding affinity, subcellular localization, and posttranslational modification including phosphorylation. We have previously identified Polo-like kinase 1(Plk1) and Casein kinase 2α (CK2 α) as the tfirs two kinases proven to phosphorylate YY1. In this study, we
Load more

Recommended publications
  • TGF-Β1 Signaling Targets Metastasis-Associated Protein 1, a New Effector in Epithelial Cells
    Oncogene (2011) 30, 2230–2241 & 2011 Macmillan Publishers Limited All rights reserved 0950-9232/11 www.nature.com/onc ORIGINAL ARTICLE TGF-b1 signaling targets metastasis-associated protein 1, a new effector in epithelial cells SB Pakala1, K Singh1,3, SDN Reddy1, K Ohshiro1, D-Q Li1, L Mishra2 and R Kumar1 1Department of Biochemistry and Molecular Biology and Institute of Coregulator Biology, The George Washington University Medical Center, Washington, DC, USA and 2Department of Gastroenterology, Hepatology and Nutrition, The University of Texas MD Anderson Cancer Center, Houston, TX, USA In spite of a large number of transforming growth factor b1 gene chromatin in response to upstream signals. The (TGF-b1)-regulated genes, the nature of its targets with TGF-b1-signaling is largely mediated by Smad proteins roles in transformation continues to be poorly understood. (Massague et al., 2005) where Smad2 and Smad3 are Here, we discovered that TGF-b1 stimulates transcription phosphorylated by TGF-b1-receptors and associate with of metastasis-associated protein 1 (MTA1), a dual master the common mediator Smad4, which translocates to the coregulator, in epithelial cells, and that MTA1 status is a nucleus to participate in the expression of TGF-b1-target determinant of TGF-b1-induced epithelial-to-mesenchymal genes (Deckers et al., 2006). Previous studies have shown transition (EMT) phenotypes. In addition, we found that that CUTL1, also known as CDP (CCAAT displacement MTA1/polymerase II/activator protein-1 (AP-1) co-activator protein), a target of TGF-b1, is needed for its short-term complex interacts with the FosB-gene chromatin and stimu- effects of TGF-b1 on cell motility involving Smad4- lates its transcription, and FosB in turn, utilizes FosB/histone dependent pathway (Michl et al.,2005).
    [Show full text]
  • Conditional Deletion of HIF-1Α Provides New Insight
    bioRxiv preprint doi: https://doi.org/10.1101/2021.01.16.426940; this version posted January 16, 2021. 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. Conditional deletion of HIF-1 provides new insight regarding the murine response to gastrointestinal infection with Salmonella Typhimurium Laura Robrahn *, Aline Dupont †, Sandra Jumpertz *, Kaiyi Zhang †, Christian H. Holland ‡ §, Joël Guillaume ¶1, Sabrina Rappold *, Vuk Cerovic ¶, Julio Saez-Rodriguez ‡ §, Mathias W. Hornef † || #, and Thorsten Cramer * ** †† ‡‡ * Department of General, Visceral and Transplantation Surgery, RWTH University Hospital, Pauwelsstraße 30, 52074 Aachen, Germany; † Institute of Medical Microbiology, RWTH University Hospital, Pauwelsstraße 30, 52074 Aachen, Germany; ‡ Institute for Computational Biomedicine, Faculty of Medicine, Heidelberg University, and Heidelberg University Hospital, Bioquant, Heidelberg, Germany; § Joint Research Center for Computational Biomedicine, RWTH Aachen University Hospital, Aachen, Germany; ¶ Institute of Molecular Medicine, RWTH University Hospital, Aachen, Germany; ** ESCAM – European Surgery Center Aachen Maastricht, Germany and The Netherlands; †† Department of Surgery, Maastricht University Medical Center, Maastricht, The Netherlands; ‡‡ NUTRIM School of Nutrition and Translational Research in Metabolism, Maastricht University, Maastricht,
    [Show full text]
  • Rbpj-Dependent and -Independent Notch2 Signaling Regulates
    RBPJ-DEPENDENT AND -INDEPENDENT NOTCH2 SIGNALING REGULATES CILIARY BODY DEVELOPMENT IN THE MOUSE EYE By Yi Zhou B.S., Tsinghua University, 2010 Submitted to the graduate degree program in Anatomy and Cell Biology and the Graduate Faculty of the University of Kansas in partial fulfillment of the requirements for the degree of Doctor of Philosophy. ________________________________ Ting Xie, Ph.D., Co-Chair ________________________________ William Kinsey, Ph.D., Co-Chair ________________________________ Dale Abrahamson, Ph.D. ________________________________ Peter Smith, Ph.D. ________________________________ Jerry Workman, Ph.D. Date Defended: Jan 6th, 2016 The Dissertation Committee for Yi Zhou certifies that this is the approved version of the following dissertation: RBPJ-DEPENDENT AND -INDEPENDENT NOTCH2 SIGNALING REGULATES CILIARY BODY DEVELOPMENT IN THE MOUSE EYE ________________________________ Ting Xie, Ph.D., Co-Chair ________________________________ William Kinsey, Ph.D., Co-Chair Date Approved: Jan 15th, 2016 ii ABSTRACT The ciliary body (CB) is a two-layered structure in the anterior eye, which is composed of the pigmented outer ciliary epithelium (OCE) and the non-pigmented inner ciliary epithelium (ICE). It is responsible for aqueous humor secretion and lens accommodation. Despite the important roles in maintaining normal eye functions, its development still remains poorly understood. The Notch signaling pathway is an evolutionarily conserved pathway that has diverse functions during tissue development and homeostasis. Canonical Notch signaling is mediated through the recombination signal binding protein for immunoglobulin kappa J region (RBPJ)-dependent transcription activation and repression. In this study, I have demonstrated that Notch2 and RBPJ are important regulators of CB development by conditionally deleting them in the developing CB.
    [Show full text]
  • Estradiol Inhibits ER Stress-Induced Apoptosis Through
    Laboratory Investigation (2014) 94, 906–916 & 2014 USCAP, Inc All rights reserved 0023-6837/14 17b-Estradiol inhibits ER stress-induced apoptosis through promotion of TFII-I-dependent Grp78 induction in osteoblasts Yun-Shan Guo1,2,5, Zhen Sun1,5, Jie Ma3,5, Wei Cui4,5, Bo Gao1, Hong-Yang Zhang1, Yue-Hu Han1, Hui-Min Hu1, Long Wang1, Jing Fan1, Liu Yang1, Juan Tang2 and Zhuo-Jing Luo1 Although many studies have suggested that estrogen prevents postmenopausal bone loss partially due to its anti-apoptosis effects in osteoblasts, the underlying mechanism has not been fully elucidated. In the present study, we found that 17b-estradiol (17b-E2), one of the primary estrogens, inhibited endoplasmic reticulum (ER) stress-induced apoptosis in MC3T3-E1 cells and primary osteoblasts. Interestingly, 17b-E2-promoted Grp78 induction, but not CHOP induction in response to ER stress. We further confirmed that Grp78-specific siRNA reversed the inhibition of 17b-E2 on ER stress-induced apoptosis by activating caspase-12 and caspase-3. Moreover, we found that 17b-E2 markedly increased the phosphorylated TFII-I levels and nuclear localization of TFII-I in ER stress conditions. 17b-E2 stimulated Grp78 promoter activity in a dose-dependent manner in the presence of TFII-I and enhanced the binding of TFII-I to the Grp78 promoter. In addition, 17b-E2 notably increased phosphorylated ERK1/2 levels and Ras kinase activity in MC3T3-E1 cells. The ERK1/2 activity-specific inhibitor U0126 remarkably blocked 17b-E2-induced TFII-I phosphorylation and Grp78 expression in response to ER stress. Together, 17b-E2 protected MC3T3-E1 cells against ER stress-induced apoptosis by promoting Ras-ERK1/2-TFII-I signaling pathway-dependent Grp78 induction.
    [Show full text]
  • In Vitro Targeting of Transcription Factors to Control the Cytokine Release Syndrome in 2 COVID-19 3
    bioRxiv preprint doi: https://doi.org/10.1101/2020.12.29.424728; this version posted December 30, 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 4.0 International license. 1 In vitro Targeting of Transcription Factors to Control the Cytokine Release Syndrome in 2 COVID-19 3 4 Clarissa S. Santoso1, Zhaorong Li2, Jaice T. Rottenberg1, Xing Liu1, Vivian X. Shen1, Juan I. 5 Fuxman Bass1,2 6 7 1Department of Biology, Boston University, Boston, MA 02215, USA; 2Bioinformatics Program, 8 Boston University, Boston, MA 02215, USA 9 10 Corresponding author: 11 Juan I. Fuxman Bass 12 Boston University 13 5 Cummington Mall 14 Boston, MA 02215 15 Email: [email protected] 16 Phone: 617-353-2448 17 18 Classification: Biological Sciences 19 20 Keywords: COVID-19, cytokine release syndrome, cytokine storm, drug repurposing, 21 transcriptional regulators 1 bioRxiv preprint doi: https://doi.org/10.1101/2020.12.29.424728; this version posted December 30, 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 4.0 International license. 22 Abstract 23 Treatment of the cytokine release syndrome (CRS) has become an important part of rescuing 24 hospitalized COVID-19 patients. Here, we systematically explored the transcriptional regulators 25 of inflammatory cytokines involved in the COVID-19 CRS to identify candidate transcription 26 factors (TFs) for therapeutic targeting using approved drugs.
    [Show full text]
  • Enhancer, Hs3 Igh ′ Activator of the Murine 3 Yin Yang 1 Is A
    Yin Yang 1 Is a Lipopolysaccharide-Inducible Activator of the Murine 3 ′ Igh Enhancer, hs3 This information is current as Steven J. Gordon, Shireen Saleque and Barbara K. Birshtein of September 23, 2021. J Immunol 2003; 170:5549-5557; ; doi: 10.4049/jimmunol.170.11.5549 http://www.jimmunol.org/content/170/11/5549 Downloaded from References This article cites 66 articles, 40 of which you can access for free at: http://www.jimmunol.org/content/170/11/5549.full#ref-list-1 Why The JI? Submit online. http://www.jimmunol.org/ • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication *average by guest on September 23, 2021 Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2003 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology Yin Yang 1 Is a Lipopolysaccharide-Inducible Activator of the Murine 3؅ Igh Enhancer, hs31 Steven J. Gordon, Shireen Saleque,2 and Barbara K. Birshtein3 The 3؅ Igh enhancers, DNase I hypersensitive site (hs) 3B and/or hs4, are required for germline transcription, and hence, class switch recombination for multiple isotypes.
    [Show full text]
  • Endothelial Specific YY1 Deletion Restricts Tumor Angiogenesis And
    www.nature.com/scientificreports OPEN Endothelial specifc YY1 deletion restricts tumor angiogenesis and tumor growth Huan Liu1,2, Yikai Qiu1, Xiuying Pei1, Ramamurthy Chitteti2, Rebbeca Steiner2, Shuya Zhang1,2* & Zheng Gen Jin2* Angiogenesis is a physiological process for the formation of new blood vessels from the pre-existing vessels and it has a vital role in the survival and growth of neoplasms. During tumor angiogenesis, the activation of the gene transcriptions in vascular endothelial cells (ECs) plays an essential role in the promotion of EC proliferation, migration, and vascular network development. However, the molecular mechanisms underlying transcriptional regulation of EC and tumor angiogenesis remains to be fully elucidated. Here we report that the transcription factor Yin Yang 1 (YY1) in ECs is critically involved in tumor angiogenesis. First, we utilized a tamoxifen-inducible EC-specifc YY1 defcient mouse model and showed that YY1 deletion in ECs inhibited the tumor growth and tumor angiogenesis. Using the in vivo matrigel plug assay, we then found that EC-specifc YY1 ablation inhibited growth factor-induced angiogenesis. Furthermore, vascular endothelial growth factor (VEGF)-induced EC migration was diminished in YY1-depleted human umbilical vein endothelial cells (HUVECs). Finally, a rescue experiment revealed that YY1-regulated BMP6 expression in ECs was involved in EC migration. Collectively, our results demonstrate that endothelial YY1 has a crucial role in tumor angiogenesis and suggest that targeting endothelial
    [Show full text]
  • The YY1 Transcription Factor Is a Component of Ribonucleoprotein Complexes In
    The YY1 Transcription Factor is a Component of Ribonucleoprotein Complexes in Xenopus laevis Oocytes and Embryos. A Thesis Submitted to the College of Graduate Studies and Research In Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy In the Department of Anatomy and Cell Biology University of Saskatchewan Saskatoon Saskatchewan By Andrew Douglas Ficzycz Spring 2003 © Copyright Andrew Do uglas Ficzyc z, 2003. All rights reserv ed. Permission to use P ERMIS SION TO US E I n presenting this thesis in partial f ulfilment of the r equirements for a P ostgraduate degree f rom the University of S askatchew an, I agree that the Libraries of this U nivers ity may make it f reely available f or inspection. I f urther agree that permission f or copying of this thesis in any manner, in w hole or in part, for s cholarly purpos es may be granted by the professor or profess ors w ho supervis ed my thesis w ork or , in their absence, by the Head of the D epartment or the D ean of the College in w hich my thesis work was done. I t is unders tood that any copying or publication or use of this thesis or parts thereof f or financial gain s hall not be allowed without my w ritten permis sion. It is also understood that due r ecognition shall be given to me and to the Univer s ity of Saskatchew an in any scholarly use w hich may be made of any material in my thesis.
    [Show full text]
  • KRAS/NF-Kb/YY1/Mir-489 Signaling Axis Controls Pancreatic Cancer
    Published OnlineFirst October 28, 2016; DOI: 10.1158/0008-5472.CAN-16-1898 Cancer Molecular and Cellular Pathobiology Research KRAS/NF-kB/YY1/miR-489 Signaling Axis Controls Pancreatic Cancer Metastasis Peng Yuan1,2, Xiao-Hong He1,2, Ye-Fei Rong3, Jing Cao4, Yong Li5, Yun-Ping Hu6, Yingbin Liu6, Dangsheng Li7, Wenhui Lou3, and Mo-Fang Liu1,2,4 Abstract KRAS activation occurring in more than 90% of pancreatic inhibited migration and metastasis by targeting the extracellular ductal adenocarcinomas (PDAC) drives progression and metas- matrix factors ADAM9 and MMP7. miR-489 downregulation tasis, but the underlying mechanisms involved in these processes elevated levels of ADAM9 and MMP7, thereby enhancing are still poorly understood. Here, we show how KRAS acts through the migration and metastasis of PDAC cells. Together, our inflammatory NF-kB signaling to activate the transcription factor results establish a pivotal mechanism of PDAC metastasis and YY1, which represses expression of the tumor suppressor gene suggest miR-489 as a candidate therapeutic target for their attack. miR-489. In PDAC cells, repression of miR-489 by KRAS signaling Cancer Res; 77(1); 100–11. Ó2016 AACR. KRAS in pancreatic tumors, the most common of which is Introduction G12D KRAS (4). Mounting evidence indicates that such onco- Pancreatic cancer (PDAC) is one of the most lethal malignant genic mutations play critical roles in both the initiation and the tumors, with a 5-year survival rate less than 8% from 2005 to 2011 progression of pancreatic cancer via persistent activation of in United States (1). To date, lack of effective screening tool to KRAS signaling pathways (5).
    [Show full text]
  • Relief of YY1 Transcriptional Repression by Adenovirus E1A Is Mediated by E1A-Associated Protein P300
    Downloaded from genesdev.cshlp.org on September 25, 2021 - Published by Cold Spring Harbor Laboratory Press Relief of YY1 transcriptional repression by adenovirus E1A is mediated by E1A-associated protein p300 Jeng-Shin Lee, 1"2 Katherine M. Galvin, l's Raymond H. See, l's Richard Eckner, 3 David Livingston, 3 Elizabeth Moran, 4 and Yang Shi 1'2'6 ~Department of Pathology, 2Committee on Virology, Harvard Medical School, Boston, Massachusetts 02115 USA; aDivision of Cellular and Molecular Biology, Dana-Farber Cancer Institute, Boston, Massachusetts 02115 USA; 4The Fels Institute for Cancer Research and Molecular Biology, Temple University School of Medicine, Philadelphia, Pennsylvania 19140 USA YY1 represses transcription when bound upstream of transcriptional initiation sites. This repression can be relieved by adenovirus E1A. Here, we present genetic evidence that the ability of E1A to relieve YY1 repression was impaired by mutations that affect EIA binding to its associated protein p300. This suggests that E1A may modulate the repressor activity of YY1 by binding to p300, which may be physically complexed with YY1. A YY1/p300 protein complex in vivo was demonstrated by several independent approaches, and the YYl-interacting domain was mapped to the carboxy-terminal region of p300, distinct from the E1A-binding site. Unlike E2F/RB, the YY1/p300 complex is not disrupted by E1A. Functional studies using recombinant p300 demonstrated unequivocally that p300 is capable of mediating E1A-induced transcriptional activation through YY1. Taken together, these results reveal, for the first time, a YY1/p300 complex that is targeted by EIA and demonstrate a function for p300 in mediating interactions between YY1 and E1A.
    [Show full text]
  • PDF-Document
    Supplementary Material Investigating the role of microRNA and Transcription Factor co-regulatory networks in Multiple Sclerosis pathogenesis Nicoletta Nuzziello1, Laura Vilardo2, Paride Pelucchi2, Arianna Consiglio1, Sabino Liuni1, Maria Trojano3 and Maria Liguori1* 1National Research Council, Institute of Biomedical Technologies, Bari Unit, Bari, Italy 2National Research Council, Institute of Biomedical Technologies, Segrate Unit, Milan, Italy 3Department of Basic Sciences, Neurosciences and Sense Organs, University of Bari, Bari, Italy Supplementary Figure S1 Frequencies of GO terms and canonical pathways. (a) Histogram illustrates the GO terms associated to assembled sub-networks. (b) Histogram illustrates the canonical pathways associated to assembled sub-network. a b Legends for Supplementary Tables Supplementary Table S1 List of feedback (FBL) and feed-forward (FFL) loops in miRNA-TF co-regulatory network. Supplementary Table S2 List of significantly (adj p-value < 0.05) GO-term involved in MS. The first column (from the left) listed the GO-term (biological processes) involved in MS. For each functional class, the main attributes (gene count, p-value, adjusted p-value of the enriched terms for multiple testing using the Benjamini correction) have been detailed. In the last column (on the right), we summarized the target genes involved in each enriched GO-term. Supplementary Table S3 List of significantly (adj p-value < 0.05) enriched pathway involved in MS. The first column (from the left) listed the enriched pathway involved in MS. For each pathway, the main attributes (gene count, p-value, adjusted p-value of the enriched terms for multiple testing using the Benjamini correction) have been detailed. In the last column (on the right), we summarized the target genes involved in each enriched pathway.
    [Show full text]
  • YY1 Suppresses FEN1 Over-Expression and Drug
    Wang et al. BMC Cancer (2015) 15:50 DOI 10.1186/s12885-015-1043-1 RESEARCH ARTICLE Open Access YY1 suppresses FEN1 over-expression and drug resistance in breast cancer Jianwei Wang1, Lina Zhou1,2, Zhi Li3, Ting Zhang1, Wenpeng Liu1, Zheng Liu2, Yate-Ching Yuan2, Fan Su2,LuXu3, Yan Wang3, Xiaotong Zhou3, Hong Xu4, Yuejin Hua4, Ying-Jie Wang5, Li Zheng2, Yue-E Teng3* and Binghui Shen2* Abstract Background: Drug resistance is a major challenge in cancer therapeutics. Abundant evidence indicates that DNA repair systems are enhanced after repetitive chemotherapeutic treatments, rendering cancers cells drug-resistant. Flap endonuclease 1 (FEN1) plays critical roles in DNA replication and repair and in counteracting replication stress, which is a key mechanism for many chemotherapeutic drugs to kill cancer cells. FEN1 was previously shown to be upregulated in response to DNA damaging agents. However, it is unclear about the transcription factors that regulate FEN1 expression in human cancer. More importantly, it is unknown whether up-regulation of FEN1 has an adverse impact on the prognosis of chemotherapeutic treatments of human cancers. Methods: To reveal regulation mechanism of FEN1 expression, we search and identify FEN1 transcription factors or repressors and investigate their function on FEN1 expression by using a combination of biochemical, molecular, and cellular approaches. Furthermore, to gain insights into the impact of FEN1 levels on the response of human cancer to therapeutic treatments, we determine FEN1 levels in human breast cancer specimens and correlate them to the response to treatments and the survivorship of corresponding breast cancer patients. Results: We observe that FEN1 is significantly up-regulated upon treatment of chemotherapeutic drugs such as mitomycin C (MMC) and Taxol in breast cancer cells.
    [Show full text]