DOCSLIB.ORG

Taz As a Regulator of Mesenchymal Transformation and Clinical Aggressiveness in Gliomas

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Taz As a Regulator of Mesenchymal Transformation and Clinical Aggressiveness in Gliomas The Texas Medical Center Library DigitalCommons@TMC The University of Texas MD Anderson Cancer Center UTHealth Graduate School of The University of Texas MD Anderson Cancer Biomedical Sciences Dissertations and Theses Center UTHealth Graduate School of (Open Access) Biomedical Sciences 5-2012 TAZ AS A REGULATOR OF MESENCHYMAL TRANSFORMATION AND CLINICAL AGGRESSIVENESS IN GLIOMAS Katrina Salazar Follow this and additional works at: https://digitalcommons.library.tmc.edu/utgsbs_dissertations Part of the Medical Molecular Biology Commons, Medical Pathology Commons, Neoplasms Commons, Oncology Commons, and the Pathology Commons Recommended Citation Salazar, Katrina, "TAZ AS A REGULATOR OF MESENCHYMAL TRANSFORMATION AND CLINICAL AGGRESSIVENESS IN GLIOMAS" (2012). The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences Dissertations and Theses (Open Access). 229. https://digitalcommons.library.tmc.edu/utgsbs_dissertations/229 This Dissertation (PhD) is brought to you for free and open access by the The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences at DigitalCommons@TMC. It has been accepted for inclusion in The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences Dissertations and Theses (Open Access) by an authorized administrator of DigitalCommons@TMC. For more information, please contact [email protected]. TAZ AS A REGULATOR OF MESENCHYMAL TRANSFORMATION AND CLINICAL AGGRESSIVENESS IN GLIOMAS by Katrina Lumen Salazar, B.S. APPROVED: ______________________________ Kenneth Aldape, M.D. Supervisory Professor ______________________________ Russell Broaddus, M.D./Ph.D. ______________________________ Daniel Cahill, M.D./Ph.D. ______________________________ Pierre McCrea, Ph.D. ______________________________ Erik Sulman, M.D/Ph.D. APPROVED: ____________________________ Dean, The University of Texas Graduate School of Biomedical Sciences at Houston TAZ AS A REGULATOR OF MESENCHYMAL TRANSFORMATION AND CLINICAL AGGRESSIVENESS IN GLIOMAS A DISSERTATION Presented to the Faculty of The University of Texas Health Science Center at Houston and The University of Texas M. D. Anderson Cancer Center Graduate School of Biomedical Sciences in Partial Fulfillment of the Requirements for the Degree of DOCTOR OF PHILOSOPHY by Katrina Lumen Salazar, B.S. Houston, Texas May 2012 COPYRIGHT iii DEDICATION This dissertation is dedicated to my mother, Sonia Lumen Salazar (1946-2006), to whom I owe my ambition, my independent nature, and my interest in medicine. iv TAZ AS A REGULATOR OF MESENCHYMAL TRANSFORMATION AND CLINICAL AGGRESSIVENESS IN GLIOMAS Publication No.________ Katrina Lumen Salazar, B.S. Supervisory Professor: Kenneth Aldape, M.D. Glioblastoma multiforme (GBM) is an aggressive, high grade brain tumor. Microarray studies have shown a subset of GBMs with a mesenchymal gene signature. This subset is associated with poor clinical outcome and resistance to treatment. To establish the molecular drivers of this mesenchymal transition, we correlated transcription factor expression to the mesenchymal signature and identified transcriptional co-activator with PDZ-binding motif (TAZ) to be highly associated with the mesenchymal shift. High TAZ expression correlated with worse clinical outcome and higher grade. These data led to the hypothesis that TAZ is critical to the mesenchymal transition and aggressive clinical behavior seen in GBM. We investigated the expression of TAZ, its binding partner TEAD, and the mesenchymal marker FN1 in human gliomas. Western analyses demonstrated increased expression of TAZ, TEAD4, and FN1 in GBM relative to lower grade gliomas. We also identified CpG islands in the TAZ promoter that are methylated in most lower grade gliomas, but not in GBMs. TAZ-methylated glioma stem cell (GSC) lines treated with a demethylation agent showed an increase in mRNA and protein TAZ expression; therefore, methylation may be another novel way TAZ is regulated since TAZ is epigenetically silenced in tumors with a better clinical outcome. v To further characterize the role of TAZ in gliomagenesis, we stably silenced or over- expressed TAZ in GSCs. Silencing of TAZ decreased invasion, self-renewal, mesenchymal protein expression, and tumor-initiating capacity. Over-expression of TAZ led to an increase in invasion, mesenchymal protein expression, mesenchymal differentiation, and tumor-initiating ability. These actions are dependent on TAZ interacting with TEAD since all these effects were abrogated with TAZ could not bind to TEAD. We also show that TAZ and TEAD directly bind to mesenchymal gene promoters. Thus, TAZ-TEAD interaction is critically important in the mesenchymal shift and in the aggressive clinical behavior of GBM. We identified TAZ as a regulator of the mesenchymal transition in gliomas. TAZ could be used as a biomarker to both estimate prognosis and stratify patients into clinically relevant subgroups. Since mesenchymal transition is correlated to tumor aggressiveness, strategies to target and inhibit TAZ-TEAD and the downstream gene targets may be warranted in alternative treatment. vi TABLE OF CONTENTS COPYRIGHT .............................................................................................................................. iii DEDICATION ............................................................................................................................. iv ABSTRACT ................................................................................................................................. v LIST OF ILLUSTRATIONS ....................................................................................................... ix LIST OF TABLES ...................................................................................................................... xii CHAPTER 1: INTRODUCTION ................................................................................................. 1 Glioblastoma multiforme (GBM) ............................................................................................. 2 The mesenchymal signature and GBM ..................................................................................... 5 The Hippo Pathway ................................................................................................................... 8 TAZ, YAP, and TEAD ........................................................................................................... 11 EMT versus PMT .................................................................................................................... 22 Hypothesis and Significance ................................................................................................... 26 CHAPTER 2: METHODS .......................................................................................................... 28 CHAPTER 3: RESULTS—SPECIFIC AIM 1 ........................................................................... 60 TAZ is associated with both the mesenchymal signature and GBM ...................................... 61 TAZ is essential for PMT and aggressive behavior in glioma stem cells (GSCs) .................. 75 CHAPTER 4: RESULTS—SPECIFIC AIM 2 ........................................................................... 88 The mesenchymal transition is controlled by TAZ interacting with TEAD ........................... 89 vii TAZ-TEAD directly binds the promoters of target genes to induce mesenchymal transition ............................................................................................................................................... 101 TAZ-TEAD interaction decreases survival and increases glioma grade in a mouse model . 110 CHAPTER 5: DISCUSSION ................................................................................................... 118 Summary ............................................................................................................................... 119 Future Directions, Clinical Implications, and Conclusions .................................................. 123 CHAPTER 6: APPENDIX ....................................................................................................... 129 WT-YAP decreases survival and increases glioma grade in the RCAS/N-tva model .......... 130 TGF-β does not activate TAZ in GSCs ................................................................................. 133 GSCs cultured with CTGF show an increase in growth ....................................................... 138 TAZ-TEAD may recruit SATB1/2 to repress proneural genes ............................................ 140 BIBLIOGRAPHY..................................................................................................................... 142 VITA ......................................................................................................................................... 200 viii LIST OF ILLUSTRATIONS Figure 1: The Hippo pathway ....................................................................................................... 9 Figure 2: Kaplan-Meier analyses of patient survival in TCGA datasets .................................... 62 Figure 3: TAZ/WWTR1 expression versus mesenchymal metagene score ................................. 64 Figure 4: The Hippo pathway (simplified) ................................................................................. 65 Figure 5: Methylation status of WWTR1 and YAP1...................................................................
Load more

Recommended publications
  • Machine-Learning and Chemicogenomics Approach Defi Nes and Predicts Cross-Talk of Hippo and MAPK Pathways
    Published OnlineFirst November 18, 2020; DOI: 10.1158/2159-8290.CD-20-0706 RESEARCH ARTICLE Machine -Learning and Chemicogenomics Approach Defi nes and Predicts Cross-Talk of Hippo and MAPK Pathways Trang H. Pham 1 , Thijs J. Hagenbeek 1 , Ho-June Lee 1 , Jason Li 2 , Christopher M. Rose 3 , Eva Lin 1 , Mamie Yu 1 , Scott E. Martin1 , Robert Piskol 2 , Jennifer A. Lacap 4 , Deepak Sampath 4 , Victoria C. Pham 3 , Zora Modrusan 5 , Jennie R. Lill3 , Christiaan Klijn 2 , Shiva Malek 1 , Matthew T. Chang 2 , and Anwesha Dey 1 ABSTRACT Hippo pathway dysregulation occurs in multiple cancers through genetic and non- genetic alterations, resulting in translocation of YAP to the nucleus and activation of the TEAD family of transcription factors. Unlike other oncogenic pathways such as RAS, defi ning tumors that are Hippo pathway–dependent is far more complex due to the lack of hotspot genetic alterations. Here, we developed a machine-learning framework to identify a robust, cancer type–agnostic gene expression signature to quantitate Hippo pathway activity and cross-talk as well as predict YAP/TEAD dependency across cancers. Further, through chemical genetic interaction screens and multiomics analyses, we discover a direct interaction between MAPK signaling and TEAD stability such that knockdown of YAP combined with MEK inhibition results in robust inhibition of tumor cell growth in Hippo dysregulated tumors. This multifaceted approach underscores how computational models combined with experimental studies can inform precision medicine approaches including predictive diagnostics and combination strategies. SIGNIFICANCE: An integrated chemicogenomics strategy was developed to identify a lineage- independent signature for the Hippo pathway in cancers.
    [Show full text]
  • Common and Distinctive Functions of the Hippo Effectors Taz and Yap In
    TISSUE-SPECIFIC STEM CELLS aRandall Division of Cell and Common and Distinctive Functions of the Hippo Molecular Biophysics, King’s College London, London, UK; Effectors Taz and Yap in Skeletal Muscle Stem Cell bSchool of Medicine, Medical Sciences & Nutrition, Function University of Aberdeen, Foresterhill, Aberdeen, a b b a Scotland, UK; cSystems CONGSHAN SUN, VANESSA DE MELLO, ABDALLA MOHAMED, HUASCAR P. O RTUSTE QUIROGA, c b d,e,f Biology Ireland, Conway AMAYA GARCIA-MUNOZ, ABDULLAH AL BLOSHI, ANNIE M. TREMBLAY, c g b c Institute, Dublin, Ireland; ALEXANDER VON KRIEGSHEIM, ELAINA COLLIE-DUGUID, NEIL VARGESSON, DAVID MATALLANAS, d b,h a Stem Cell Program, HENNING WACKERHAGE, PETER S. ZAMMIT Children’s Hospital, Boston, Massachusetts, USA; Key Words. Taz • Yap • Tead • Satellite cells • Muscle stem cells eDepartment of Stem Cell and Regenerative Biology, Harvard University, ABSTRACT Cambridge, Massachusetts, USA; fHarvard Stem Cell Hippo pathway downstream effectors Yap and Taz play key roles in cell proliferation and regen- Institute, Cambridge, eration, regulating gene expression especially via Tead transcription factors. To investigate their Massachusetts, USA; gCentre role in skeletal muscle stem cells, we analyzed Taz in vivo and ex vivo in comparison with Yap. for Genome Enabled Biology Small interfering RNA knockdown or retroviral-mediated expression of wild-type human or con- and Medicine, School of stitutively active TAZ mutants in satellite cells showed that TAZ promoted proliferation, a func- Medicine, Medical Sciences tion shared with YAP. However, at later stages of myogenesis, TAZ also enhanced myogenic and Nutrition, University of differentiation of myoblasts, whereas YAP inhibits such differentiation. Functionally, while mus- Aberdeen, Foresterhill, cle growth was mildly affected in Taz (gene Wwtr1–/–) knockout mice, there were no overt Aberdeen, Scotland, UK; effects on regeneration.
    [Show full text]
  • DNA-Damage Induced Cell Death in Yap1;Wwtr1 Mutant Epidermal Basal Cells
    bioRxiv preprint doi: https://doi.org/10.1101/2021.07.23.453490; this version posted July 23, 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 4.0 International license. 1 DNA-damage induced cell death in yap1;wwtr1 mutant epidermal basal cells 2 Jason K. H. Lai1,*, Pearlyn J. Y. Toh1, Hamizah A. Cognart1, Geetika Chouhan2 & Timothy 3 E. Saunders1,3,4,5,* 4 1 Mechanobiology Institute, National University of Singapore, Singapore 117411 5 2 Department of Biological Sciences, Tata Institute of Fundamental Research, Colaba, 6 Mumbai, India 7 3 Department of Biological Sciences, National University of Singapore, Singapore 117558 8 4 Institute of Molecular and Cell Biology, A*Star, Singapore 9 5 Warwick Medical School, University of Warwick, United Kingdom 10 * Corresponding authors: J. K. H. L. ([email protected]), T. E. S. 11 ([email protected]) 12 1 bioRxiv preprint doi: https://doi.org/10.1101/2021.07.23.453490; this version posted July 23, 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 4.0 International license. 13 ABSTRACT 14 In a previous study, it was reported that Yap1 and Wwtr1 in zebrafish regulates the 15 morphogenesis of the posterior body and epidermal fin fold (Kimelman, D., et al.
    [Show full text]
  • Whole-Exome Sequencing of Metastatic Cancer and Biomarkers of Treatment Response
    Supplementary Online Content Beltran H, Eng K, Mosquera JM, et al. Whole-exome sequencing of metastatic cancer and biomarkers of treatment response. JAMA Oncol. Published online May 28, 2015. doi:10.1001/jamaoncol.2015.1313 eMethods eFigure 1. A schematic of the IPM Computational Pipeline eFigure 2. Tumor purity analysis eFigure 3. Tumor purity estimates from Pathology team versus computationally (CLONET) estimated tumor purities values for frozen tumor specimens (Spearman correlation 0.2765327, p- value = 0.03561) eFigure 4. Sequencing metrics Fresh/frozen vs. FFPE tissue eFigure 5. Somatic copy number alteration profiles by tumor type at cytogenetic map location resolution; for each cytogenetic map location the mean genes aberration frequency is reported eFigure 6. The 20 most frequently aberrant genes with respect to copy number gains/losses detected per tumor type eFigure 7. Top 50 genes with focal and large scale copy number gains (A) and losses (B) across the cohort eFigure 8. Summary of total number of copy number alterations across PM tumors eFigure 9. An example of tumor evolution looking at serial biopsies from PM222, a patient with metastatic bladder carcinoma eFigure 10. PM12 somatic mutations by coverage and allele frequency (A) and (B) mutation correlation between primary (y- axis) and brain metastasis (x-axis) eFigure 11. Point mutations across 5 metastatic sites of a 55 year old patient with metastatic prostate cancer at time of rapid autopsy eFigure 12. CT scans from patient PM137, a patient with recurrent platinum refractory metastatic urothelial carcinoma eFigure 13. Tracking tumor genomics between primary and metastatic samples from patient PM12 eFigure 14.
    [Show full text]
  • Systematic Morphological Profiling of Human Gene and Allele Function Via
    TOOLS AND RESOURCES Systematic morphological profiling of human gene and allele function via Cell Painting Mohammad Hossein Rohban1, Shantanu Singh1, Xiaoyun Wu1, Julia B Berthet2, Mark-Anthony Bray1†, Yashaswi Shrestha1, Xaralabos Varelas2, Jesse S Boehm1, Anne E Carpenter1* 1Broad Institute of MIT and Harvard, Cambridge, United States; 2Department of Biochemistry, Boston University School of Medicine, Boston, United States Abstract We hypothesized that human genes and disease-associated alleles might be systematically functionally annotated using morphological profiling of cDNA constructs, via a microscopy-based Cell Painting assay. Indeed, 50% of the 220 tested genes yielded detectable morphological profiles, which grouped into biologically meaningful gene clusters consistent with known functional annotation (e.g., the RAS-RAF-MEK-ERK cascade). We used novel subpopulation- based visualization methods to interpret the morphological changes for specific clusters. This unbiased morphologic map of gene function revealed TRAF2/c-REL negative regulation of YAP1/ WWTR1-responsive pathways. We confirmed this discovery of functional connectivity between the NF-kB pathway and Hippo pathway effectors at the transcriptional level, thereby expanding knowledge of these two signaling pathways that critically regulate tumor initiation and progression. We make the images and raw data publicly available, providing an initial morphological map of *For correspondence: anne@ major biological pathways for future study. broadinstitute.org DOI: 10.7554/eLife.24060.001 Present address: †Novartis Institutes for BioMedical Research, Cambridge, United States Introduction Competing interests: The The dramatic increase in human genome sequence data has created a significant bottleneck. The authors declare that no number of genes and variants known to be associated with most human diseases has increased dra- competing interests exist.
    [Show full text]
  • Lentivirus-Mediated RNA Interference Targeting WWTR1 in Human Colorectal Cancer Cells Inhibits Cell Proliferation in Vitro and Tumor Growth in Vivo
    ONCOLOGY REPORTS 28: 179-185, 2012 Lentivirus-mediated RNA interference targeting WWTR1 in human colorectal cancer cells inhibits cell proliferation in vitro and tumor growth in vivo JIE PAN, SHAOTANG LI, PAN CHI, ZONGBIN XU, XINGRONG LU and YING HUANG Department of Colorectal Surgery, Affiliated Union Hospital of Fujian Medical University, Fuzhou, Fujian, P.R. China Received December 2, 2011; Accepted February 10, 2012 DOI: 10.3892/or.2012.1751 Abstract. WW domain-containing transcription regulator 1 identification of novel functional molecules, knowledge of their (WWTR1) was initially identified as a transcriptional coacti- mechanisms of action and strategies for intervention (2). vator involved in the differentiation of stem cells as well as the WW-domain containing transcription regulator 1 development of multiple organs. Recently, WWTR1 has also (WWTR1), also called TAZ (transcriptional co-activator with been identified as a major component of the novel Hippo signal- PDZ binding motif), activates many transcriptional factors ling pathway important for the development of breast and lung and has important roles in the development of various tissue cancer. Here, we show for the first time that WWTR1 has an in mammals (3). WWTR1 has also been shown to regulate oncogenic function in colorectal cancer cell lines. Knockdown stem cell differentiation and self-renewal through binding with of WWTR1 by lentivirus-mediated RNA interference in human the transcription factors PPARγ, Runx2 and Smad (4,5). Most colorectal cancer cells significantly decreased cell prolifer­ recently, enhanced expression of WWTR1 has been found in ation and the colony formation of RKO cells in vitro and tumor both breast and lung cancer cell lines (6,7).
    [Show full text]
  • Regulation of Posterior Body and Epidermal Morphogenesis In
    RESEARCH ARTICLE Regulation of posterior body and epidermal morphogenesis in zebrafish by localized Yap1 and Wwtr1 David Kimelman1*, Natalie L Smith1†, Jason Kuan Han Lai2†, Didier YR Stainier2 1Department of Biochemistry, University of Washington, Seattle, United States; 2Department of Developmental Genetics, Max Planck Institute for Heart and Lung Research, Bad Nauheim, Germany Abstract The vertebrate embryo undergoes a series of dramatic morphological changes as the body extends to form the complete anterior-posterior axis during the somite-forming stages. The molecular mechanisms regulating these complex processes are still largely unknown. We show that the Hippo pathway transcriptional coactivators Yap1 and Wwtr1 are specifically localized to the presumptive epidermis and notochord, and play a critical and unexpected role in posterior body extension by regulating Fibronectin assembly underneath the presumptive epidermis and surrounding the notochord. We further find that Yap1 and Wwtr1, also via Fibronectin, have an essential role in the epidermal morphogenesis necessary to form the initial dorsal and ventral fins, a process previously thought to involve bending of an epithelial sheet, but which we now show involves concerted active cell movement. Our results reveal how the Hippo pathway transcriptional program, localized to two specific tissues, acts to control essential morphological events in the vertebrate embryo. DOI: https://doi.org/10.7554/eLife.31065.001 *For correspondence: [email protected] Introduction †These authors contributed A key step in the development of the vertebrate embryonic body is the change from the roughly equally to this work spherical-shaped embryo at the end of gastrulation to the elongated body present at the end of the somite-forming stages when the initial anterior-posterior body plan is fully established (reviewed in Competing interest: See Be´naze´raf and Pourquie´, 2013; Henrique et al., 2015; Kimelman, 2016; Wilson et al., 2009).
    [Show full text]
  • TAZ (WWTR1) (NM 015472) Human Mass Spec Standard Product Data
    OriGene Technologies, Inc. 9620 Medical Center Drive, Ste 200 Rockville, MD 20850, US Phone: +1-888-267-4436 [email protected] EU: [email protected] CN: [email protected] Product datasheet for PH304082 TAZ (WWTR1) (NM_015472) Human Mass Spec Standard Product data: Product Type: Mass Spec Standards Description: WWTR1 MS Standard C13 and N15-labeled recombinant protein (NP_056287) Species: Human Expression Host: HEK293 Expression cDNA Clone RC204082 or AA Sequence: Predicted MW: 44.1 kDa Protein Sequence: >RC204082 protein sequence Red=Cloning site Green=Tags(s) MNPASAPPPLPPPGQQVIHVTQDLDTDLEALFNSVMNPKPSSWRKKILPESFFKEPDSGSHSRQSSTDSS GGHPGPRLAGGAQHVRSHSSPASLQLGTGAGAAGSPAQQHAHLRQQSYDVTDELPLPPGWEMTFTATGQR YFLNHIEKITTWQDPRKAMNQPLNHMNLHPAVSSTPVPQRSMAVSQPNLVMNHQHQQQMAPSTLSQQNHP TQNPPAGLMSMPNALTTQQQQQQKLRLQRIQMERERIRMRQEELMRQEAALCRQLPMEAETLAPVQAAVN PPTMTPDMRSITNNSSDPFLNGGPYHSREQSTDSGLGLGCYSVPTTPEDFLSNVDEMDTGENAGQTPMNI NPQQTRFPDFLDCLPGTNVDLGTLESEDLIPLFNDVESALNKSEPFLTWL TRTRPLEQKLISEEDLAANDILDYKDDDDKV Tag: C-Myc/DDK Purity: > 80% as determined by SDS-PAGE and Coomassie blue staining Concentration: 50 ug/ml as determined by BCA Labeling Method: Labeled with [U- 13C6, 15N4]-L-Arginine and [U- 13C6, 15N2]-L-Lysine Buffer: 100 mM glycine, 25 mM Tris-HCl, pH 7.3. Store at -80°C. Avoid repeated freeze-thaw cycles. Stable for 3 months from receipt of products under proper storage and handling conditions. RefSeq: NP_056287 RefSeq Size: 5135 RefSeq ORF: 1200 Synonyms: TAZ Locus ID: 25937 UniProt ID: Q9GZV5 This product is to be used for laboratory
    [Show full text]
  • Glomerulocystic Kidney Disease in Mice with a Targeted Inactivation of Wwtr1
    Glomerulocystic kidney disease in mice with a targeted inactivation of Wwtr1 Zakir Hossain*†, Safiah Mohamed Ali*, Hui Ling Ko*, Jianliang Xu*, Chee Peng Ng‡, Ke Guo§, Zeng Qi§, Sathivel Ponniah*, Wanjin Hong‡, and Walter Hunziker*¶ *Epithelial Cell Biology Laboratory, ‡Membrane Biology Laboratory, and §Histology Unit, Institute of Molecular and Cell Biology, 61 Biopolis Drive, Republic of Singapore 138673 Edited by Christine E. Seidman, Harvard Medical School, Boston, MA, and approved November 29, 2006 (received for review June 22, 2006) Wwtr1 is a widely expressed 14-3-3-binding protein that regulates An increasing number of genes linked to PKD have been the activity of several transcription factors involved in develop- shown to encode proteins associated with the structure and/or ment and disease. To elucidate the physiological role of Wwtr1, we function of primary cilia, strongly suggesting that defects in the .(generated Wwtr1؊/؊ mice by homologous recombination. Surpris- ciliary apparatus play a central role in the etiology of PKD (4–6 ingly, although Wwtr1 is known to regulate the activity of Cbfa1, Primary or nonmotile cilia are structurally related to motile a transcription factor important for bone development, Wwtr1؊/؊ flagella of sperm and protozoa but serve as mechano- or mice show only minor skeletal defects. However, Wwtr1؊/؊ ani- chemosensors (reviewed in ref. 4). Cilia are anchored to the basal mals present with renal cysts that lead to end-stage renal disease. body, and their structural unit, the axoneme, consists of micro- Cysts predominantly originate from the dilation of Bowman’s tubules, dynein arms, and radial spoke proteins. Cilia assembly spaces and atrophy of glomerular tufts, reminiscent of glomeru- and maintenance involves the antero- and retrograde transport locystic kidney disease in humans.
    [Show full text]
  • Original Article LATS1 Inhibits Metastasis and Epithelial-Mesenchymal Transition in Head and Neck Squamous Cell Carcinoma
    Int J Clin Exp Pathol 2018;11(4):2053-2063 www.ijcep.com /ISSN:1936-2625/IJCEP0073054 Original Article LATS1 inhibits metastasis and epithelial-mesenchymal transition in head and neck squamous cell carcinoma Jicheng Wu1, Zhijuan Zhao2, Huiling Zhang1, Fanshuang Kong1, Huamao Jiang3, Keqiang Huang4, Huachuan Zheng1 1Department of Experimental Oncology, Shengjing Hospital of China Medical University, Shenyang, China; Depart- ments of 2Pathology, 3Urology, The First Affiliated Hospital of Jinzhou Medical University, Jinzhou, China; 4Depart- ment of Stomatology, The Second Affiliated Hospital of Jinzhou Medical University, Jinzhou, China Received January 19, 2018; Accepted February 22, 2018; Epub April 1, 2018; Published April 15, 2018 Abstract: LATS1 is a serine/threonine kinase of the Hippo signaling pathway that phosphorylates and inactivates transcriptional co-activators YAP1 and WWTR1. To investigate roles of LATS1 expression in head and neck squamous cell carcinomas (HNSCCs), we transfected LATS1-expressing plasmid into B88 cells and examined the phenotypes and their relevant molecules. LATS1 expression was analyzed using immunohistochemistry on tissue microarray, Oncomine, and TCGA databases. LATS1 overexpression was found to suppress growth, migration and invasion, and induce apoptosis, G2 arrest, and mesenchymal to epithelial transition (MET) (P < 0.05). Both increased expression of P21, Bax, and E-cadherin and decreased expression of Cyclin B1, D1, Bcl-2, and MMPs. Twist and N-cadherin were detected in B88 transfectants, in comparison to mock and control by Western blot. Nuclear LATS1 expression was weaker in primary cancers than in normal squamous tissue and dysplasia (P < 0.05) but versa for cytoplasmic counterpart (P < 0.05). Cytoplasmic LATS1 expression was positively correlated with lymph node metastasis (P < 0.05).
    [Show full text]
  • Phospho-STK4 Thr183 Polyclonal Antibody
    For Research Use Only Phospho-MST1 (Thr183) Polyclonal antibody www.ptglab.com Catalog Number:28953-1-AP Catalog Number: GenBank Accession Number: Purification Method: Basic Information 28953-1-AP BC005231 Antigen affinity purification Size: GeneID (NCBI): Recommended Dilutions: 100ul , Concentration: 400 μg/ml by 6789 WB 1:500-1:2000 Nanodrop; Full Name: Source: serine/threonine kinase 4 Rabbit Calculated MW: Isotype: 56 kDa IgG Observed MW: 52-56 kDa Applications Tested Applications: Positive Controls: WB,ELISA WB : Staurosporine treated Ramos cells, Species Specificity: Human Mammalian STE20-like serine-threonine kinase MST1, encoded by the STK4 gene, is a multifunctional protein. MST1 Background Information and its closest paralogs MST2 (encoded by the STK3 gene), MST3, and MST4 are members of the Class II Germinal Center Family of Protein Kinases . STK3/4 and LATS1/2 (large tumor suppressor 1 and 2) are core kinase components of the Hippo tumor suppressor pathway in mammalians . In the conventional Hippo pathway, the STK3/4 and LATS1/2 signaling cascade phosphorylates and inactivates the transcriptional coactivator YAP1 (yes associated protein 1) and its close paralog WWTR1]. YAP1 and WWTR1 do not have DNA binding domains and they exert their biological outputs, such as cell proliferation and survival, by interacting with the TEAD1-4 transcription factors. Lines of evidence have indicated that dysregulation or loss of STK4/Hippo signaling is linked to developmental disorders and carcinogenesis with poor prognosis. STK4 is a stress-induced kinase and it can be activated in response to cell-death inducers. Autophosphorylation of STK4 at Thr183 (Thr180 in STK3) in the activation loop is a key activation mechanism for STK4/3 because phosphorylation of Thr183/180 causes the cleavage of STK4 by caspases under apoptotic conditions.
    [Show full text]
  • Human WWTR1 / TAZ (1-400, His-Tag) - Purified
    OriGene Technologies, Inc. OriGene Technologies GmbH 9620 Medical Center Drive, Ste 200 Schillerstr. 5 Rockville, MD 20850 32052 Herford UNITED STATES GERMANY Phone: +1-888-267-4436 Phone: +49-5221-34606-0 Fax: +1-301-340-8606 Fax: +49-5221-34606-11 [email protected] [email protected] AR51300PU-N Human WWTR1 / TAZ (1-400, His-tag) - Purified Alternate names: Transcriptional coactivator with PDZ-binding motif, WW domain-containing transcription regulator protein 1 Quantity: 0.5 mg Concentration: 0.5 mg/ml (determined by Bradford assay) Background: WWTR1 is a transcriptional coactivator which acts as a downstream regulatory target in the Hippo signaling pathway that plays a pivotal role in organ size control and tumor suppression by restricting proliferation and promoting apoptosis. The core of this pathway is composed of a kinase cascade wherein STK3/MST2 and STK4/MST1, in complex with its regulatory protein SAV1, phosphorylates and activates LATS1/2 in complex with its regulatory protein MOB1, which in turn phosphorylates and inactivates YAP1 oncoprotein and WWTR1/TAZ. Uniprot ID: Q9GZV5 NCBI: NP_001161752 GeneID: 25937 Species: Human Source: E. coli Format: State: Liquid purified protein Purity: >85% by SDS - PAGE Buffer System: 20 mM Tris-HCl buffer (pH 8.0) containing 0.4M UREA, 10% glycerol Description: Recombinant human WWTR1 protein, fused to His-tag at N-terminus, was expressed in E.coli . AA Sequence: MGSSHHHHHH SSGLVPRGSH MGSMNPASAP PPLPPPGQQV IHVTQDLDTD LEALFNSVMN PKPSSWRKKI LPESFFKEPD SGSHSRQSST DSSGGHPGPR LAGGAQHVRS HSSPASLQLG TGAGAAGSPA QQHAHLRQQS YDVTDELPLP PGWEMTFTAT GQRYFLNHIE KITTWQDPRK AMNQPLNHMN LHPAVSSTPV PQRSMAVSQP NLVMNHQHQQ QMAPSTLSQQ NHPTQNPPAG LMSMPNALTT QQQQQQKLRL QRIQMERERI RMRQEELMRQ EAALCRQLPM EAETLAPVQA AVNPPTMTPD MRSITNNSSD PFLNGGPYHS REQSTDSGLG LGCYSVPTTP EDFLSNVDEM DTGENAGQTP MNINPQQTRF PDFLDCLPGT NVDLGTLESE DLIPLFNDVE SALNKSEPFL TWL Molecular weight: 46.5 kDa (423aa) Storage: Store undiluted at 2-8°C for one week or (in aliquots) at -20°C to -80°C for longer.
    [Show full text]