DOCSLIB.ORG

Targeting Mlst8 in Mtorc2-Dependent Cancers by Laura Chonghae Kim Dissertation Submitted to the Faculty of the Graduate School

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Targeting Mlst8 in Mtorc2-Dependent Cancers by Laura Chonghae Kim Dissertation Submitted to the Faculty of the Graduate School Targeting mLST8 in mTORC2-dependent cancers By Laura Chonghae Kim Dissertation Submitted to the Faculty of the Graduate School of Vanderbilt University in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY in Cancer Biology June 30th, 2020 Nashville, Tennessee Approved: Albert B. Reynolds, Ph.D. (chair) Mark R. Boothby, M.D., Ph.D. Christine M. Lovly, M.D. Ph.D. William P. Tansey, Ph.D. Jin Chen, M.D., Ph.D. (adviser) DEDICATION This dissertation is dedicated to all cancer patients and their loved ones for their strength and courage in the fight against this disease. This is also dedicated to my family (Kalhee and Jenny Kim, Stephanie Kim) for their unwavering love and support. ii ACKNOWLEDGEMENTS Completion of this work would have been impossible without the support of many, and I would like to express my sincere gratitude here. First and foremost, I must thank my mentor, Jin Chen. Her support, mentorship, and high-fives have been invaluable in my development as a scientist, and I plan to continue seeking her guidance even when I am many years beyond graduate school. I would also like to acknowledge my committee members Al Reynolds, Bill Tansey, Mark Boothby, Rebecca Cook, and Christine Lovly, who continuously challenged me and provided scientific guidance. Much of this work stems from their insightful questions, and I am extremely appreciative of the time they put into thinking about my projects. I would also like to thank the past and present members of the Chen Lab: Shan Wang, Deanna Edwards, Wenqiang Song, Dana Brantley-Sieders, Katherine Hastings, Victoria Youngblood, Tammy Sobolik, Yoonha Hwang, Eileen Shiuan, Kalin Wilson, Verra Ngwa, Ashwin Inala, and Chris Rhee. I am tremendously grateful for their training, scientific brilliance, and most importantly their friendship throughout this journey. In particular, I would like to highlight Deanna, Shan, Wenqiang, and Eileen, who were always willing to lend an ear to my questions and musings for the last five years. I am going to miss our morning coffee runs, breakroom lunches, and even the long weekends in lab! I also need to thank those who supported me financially throughout my graduate career, specifically Dr. Jin Chen for supporting me through the Microenvironmental Influences in Cancer Training Grant and the National Cancer Institute for the Predoctoral Ruth L. Kirschstein National Research Service Award (F31). I would also like to thank all the members of the Program in Cancer Biology for day-to-day logistical support. iii Last, I must thank my friends and family. Meeting such wonderful people around Nashville and in graduate school at Vanderbilt has made this process infinitely easier. I am incredibly grateful for the wonderful memories and enduring friendship. I also feel immensely lucky to have friends from back home in North Carolina that always supported me from afar. From weekend trips to incessant messages, they all put in an incredible amount of energy into staying connected despite the distance. I cannot thank them enough for this effort. Finally, I must thank my parents, Kalhee and Jenny Kim, and sister, Stephanie, for their unconditional love and trust. Their support gave me the confidence to persevere against every hurdle, and I feel extraordinarily privileged to have them in my corner. iv TABLE OF CONTENTS DEDICATION.............................................................................................................................. ii ACKNOWLEDGEMENTS .......................................................................................................... iii LIST OF TABLES .................................................................................................................... viii LIST OF FIGURES .................................................................................................................... ix LIST OF ABBREVIATIONS ....................................................................................................... xi CHAPTERS I. Introduction .............................................................................................................. 1 Overview .............................................................................................................. 1 Lung Cancer ........................................................................................................ 2 Targeted Therapy ..................................................................................... 3 Immunotherapy ........................................................................................ 7 The mTOR Complexes ........................................................................................ 9 mTORC1 Signaling ................................................................................ 14 mTORC1 in Cancer ................................................................................ 16 mTORC2 signaling ................................................................................. 19 mTORC2 in Cancer ................................................................................ 21 mTOR Inhibitors for Cancer Therapy ...................................................... 21 mTOR in Vasculature ............................................................................. 26 mTOR in Tumor Immunity ...................................................................... 38 Summary and Thesis Projects ........................................................................... 30 II. Disruption of the scaffolding function of mLST8 selectively inhibits mTORC2 assembly and function and suppresses mTORC2-dependent tumor growth in vivo ......................................................................................................................... 33 Abstract ............................................................................................................. 33 Significance ....................................................................................................... 33 Introduction ........................................................................................................ 34 Materials and Methods ....................................................................................... 34 Cell Lines and cell culture ....................................................................... 34 Generation of the mTOR E2285A knock-in cell line by CRISPR-Cas9 technology .............................................................................................. 35 Xenograft Assay ..................................................................................... 35 Plasmids................................................................................................. 36 Lentivirus production and transduction ................................................... 36 Western Blot and Co-immunoprecipitation .............................................. 36 In Vitro Kinase Assay ............................................................................. 37 Results ............................................................................................................... 39 v mLST8 is required for mTORC2, but not mTORC1, integrity and function ............................................................................................................... 39 mLST8 mutations that uncouple binding to LBE domain of mTOR disrupt mTORC2 complex formation and signaling ............................................ 42 mLST8 serves as a molecular bridge between mTOR and SIN1 ............ 46 mLST8 point mutations that block mLST8-mTOR interactions impair mTORC2 signaling and tumor growth in vivo .......................................... 50 Discussion ......................................................................................................... 54 III. Rictor promotes NSCLC cell proliferation through formation and activation of mTORC2 at the expense of mTORC1 ................................................................... 56 Abstract ............................................................................................................. 56 Introduction ........................................................................................................ 57 Results ............................................................................................................... 58 RICTOR is amplified and overexpressed in non-small cell lung cancer .. 58 Alterations in Rictor expression lead to corresponding changes in mTORC2 and mTORC1 ......................................................................... 64 Rictor promotes proliferation of NSCLC cells .......................................... 67 Increased Rictor expression promotes NSCLC tumor growth in vivo ...... 68 Targeting mTORC2 for loss-of-function inhibits tumor growth of RICTOR- amplified NSCLC .................................................................................... 70 Discussion ......................................................................................................... 73 Materials and Methods ....................................................................................... 75 Cell lines and cell culture ........................................................................ 75 Plasmids and sgRNA Sequences ........................................................... 76 Lentivirus Production and Transduction .................................................. 76 Xenograft Assay ....................................................................................
Load more

Recommended publications
  • A Computational Approach for Defining a Signature of Β-Cell Golgi Stress in Diabetes Mellitus
    Page 1 of 781 Diabetes A Computational Approach for Defining a Signature of β-Cell Golgi Stress in Diabetes Mellitus Robert N. Bone1,6,7, Olufunmilola Oyebamiji2, Sayali Talware2, Sharmila Selvaraj2, Preethi Krishnan3,6, Farooq Syed1,6,7, Huanmei Wu2, Carmella Evans-Molina 1,3,4,5,6,7,8* Departments of 1Pediatrics, 3Medicine, 4Anatomy, Cell Biology & Physiology, 5Biochemistry & Molecular Biology, the 6Center for Diabetes & Metabolic Diseases, and the 7Herman B. Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202; 2Department of BioHealth Informatics, Indiana University-Purdue University Indianapolis, Indianapolis, IN, 46202; 8Roudebush VA Medical Center, Indianapolis, IN 46202. *Corresponding Author(s): Carmella Evans-Molina, MD, PhD ([email protected]) Indiana University School of Medicine, 635 Barnhill Drive, MS 2031A, Indianapolis, IN 46202, Telephone: (317) 274-4145, Fax (317) 274-4107 Running Title: Golgi Stress Response in Diabetes Word Count: 4358 Number of Figures: 6 Keywords: Golgi apparatus stress, Islets, β cell, Type 1 diabetes, Type 2 diabetes 1 Diabetes Publish Ahead of Print, published online August 20, 2020 Diabetes Page 2 of 781 ABSTRACT The Golgi apparatus (GA) is an important site of insulin processing and granule maturation, but whether GA organelle dysfunction and GA stress are present in the diabetic β-cell has not been tested. We utilized an informatics-based approach to develop a transcriptional signature of β-cell GA stress using existing RNA sequencing and microarray datasets generated using human islets from donors with diabetes and islets where type 1(T1D) and type 2 diabetes (T2D) had been modeled ex vivo. To narrow our results to GA-specific genes, we applied a filter set of 1,030 genes accepted as GA associated.
    [Show full text]
  • Identification of Potential Proteases for Abdominal Aortic Aneurysm by Weighted Gene Coexpression Network Analysis
    Genome Identification of potential proteases for abdominal aortic aneurysm by weighted gene coexpression network analysis Journal: Genome Manuscript ID gen-2020-0041.R1 Manuscript Type: Article Date Submitted by the 28-Jun-2020 Author: Complete List of Authors: Zhang, Hui; Peking Union Medical College Hospital, Department of Vascular Surgery Yang, Dan; Chinese Academy of Medical Sciences and Peking Union Medical College, Department of Computational Biology and Bioinformatics,Draft Institute of Medicinal Plant Development Chen, Siliang; Peking Union Medical College Hospital, Department of Vascular Surgery Li, Fangda; Peking Union Medical College Hospital, Department of Vascular Surgery Cui, Liqiang; Peking Union Medical College Hospital, Department of Vascular Surgery Liu, Zhili; Peking Union Medical College Hospital, Department of Vascular Surgery Shao, Jiang; Peking Union Medical College Hospital, Department of Vascular Surgery Chen, Yuexin; Peking Union Medical College Hospital, Department of Vascular Surgery Liu, Bao; Peking Union Medical College Hospital, Department of Vascular Surgery Zheng, Yuehong; Peking Union Medical College Hospital, Department of Vascular Surgery Abdominal aortic aneurysm, next-generation sequencing, WGCNA, Keyword: proteases, matrix metalloproteinase Is the invited manuscript for consideration in a Special Not applicable (regular submission) Issue? : https://mc06.manuscriptcentral.com/genome-pubs Page 1 of 35 Genome 1 Identification of potential proteases for abdominal aortic aneurysm by weighted gene 2 coexpression network analysis 3 Short title: WGCNA identifies crucial proteases in AAA 4 5 Hui Zhang1, Dan Yang2, Siliang Chen1, Fangda Li1, Liqiang Cui1, Zhili Liu1, Jiang Shao1, Yuexin 6 Chen1, Bao Liu1, Yuehong Zheng1. 7 1Department of Vascular Surgery, Peking Union Medical College Hospital, Beijing 100730, PR 8 China; 2Department of Computational Biology and Bioinformatics, Institute of Medicinal Plant 9 Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 10 100730, PR China.
    [Show full text]
  • Genetics of Tuberous Sclerosis Complex: Implications for Clinical Practice
    Journal name: The Application of Clinical Genetics Article Designation: REVIEW Year: 2017 Volume: 10 The Application of Clinical Genetics Dovepress Running head verso: Caban et al Running head recto: Genetics of TSC open access to scientific and medical research DOI: http://dx.doi.org/10.2147/TACG.S90262 Open Access Full Text Article REVIEW Genetics of tuberous sclerosis complex: implications for clinical practice Carolina Caban1,2 Abstract: Tuberous sclerosis complex (TSC) is a multisystem disorder that results from hetero- Nubaira Khan1,2 zygous mutations in either TSC1 or TSC2. The primary organ systems that are affected include Daphne M Hasbani3 the brain, skin, lung, kidney, and heart, all with variable frequency, penetrance, and severity. Peter B Crino1,2 Neurological features include epilepsy, autism, and intellectual disability. There are more than 1,500 known pathogenic variants for TSC1 and TSC2, including deletion, nonsense, and missense 1Department of Neurology, 2Shriners Hospitals Pediatric Research mutations, and all pathogenic mutations are inactivating, leading to loss of function effects on Center, Temple University School of the encoded proteins TSC1 and TSC2. These proteins form a complex to constitutively inhibit 3 Medicine, Department of Neurology, mechanistic target of rapamycin (mTOR) signaling cascade, and as a consequence, mTOR signal- St. Christopher’s Hospital for Children, Drexel University College ing is constitutively active within all TSC-associated lesions. The mTOR inhibitors rapamycin For personal use only. of Medicine, Philadelphia, PA, USA (sirolimus) and everolimus have been shown to reduce the size of renal and brain lesions and improve pulmonary function in TSC, and these compounds may also decrease seizure frequency.
    [Show full text]
  • Molecular Docking and Pharmacokinetic of Highly Specific Novel Pan-Mtor Inhibitors Against Solid Tumors
    MOJ Proteomics & Bioinformatics Research Article Open Access Molecular docking and pharmacokinetic of highly specific novel pan-mtor inhibitors against solid tumors Abstract Volume 5 Issue 6 - 2017 Mechanistic/mammalian target of rapamycin (mTOR) a serine/threonine kinase belonging Muhammad Naveed,1,2 Safia Zia,1 Maryam to PI3K/Akt/mTOR pathway is involved in different cellular functions cell survival, 1 1 1 metabolism, growth, proliferation, apoptosis and autophagy. Pan-mTOR inhibitors are Akhtar, Fatima Ashraf, Amber Afroz 1Department of Biochemistry and Biotechnology, University of targeted towards mTOR dysregulation, inhibiting the kinase domain of both mTORC1 Gujrat, Pakistan and mTORC2. The present study analyzes the binding modes and molecular interactions 2Department of Biotechnology, University of Gujrat, Pakistan of highly specific mTOR inhibitors, AZD8055 and its sister compoundAZD2014using computational approach. Both inhibitors proved to be effective against solid tumors in Correspondence: Muhammad Naveed, Department of vitro and in vivo. Docking analysis was performed using Auto Dock Vina, conformations Biochemistry and Biotechnology, University of Gujrat, Pakistan, were scored based upon their binding energy (kcal/mol) and illustrated using Discovery Tel 00923015524624, Email [email protected] Studio Visualizer 4.5 version. Inhibitors fit between N- and C-lobes of mTOR kinase domain into the inner hydrophobic core. The results indicated interactions with distinctive Received: February 03, 2017 | Published: June 27, 2017 mTOR residues Trp-2239, Leu-2185 and newly developed interactions with Asp-2375 for AZD2014 and with Ala-2248, His-2247, Thr-2245 for AZD8055. The binding pattern of both inhibitors was slightly different, responsible for better pharmacokinetic profile of AZD2014 and 5 fold increase in efficacy of AZD8055.
    [Show full text]
  • Structural and Functional Dissection of Reovirus Capsid Folding and Assembly by the Prefoldin-Tric/CCT Chaperone Network
    Structural and functional dissection of reovirus capsid folding and assembly by the prefoldin-TRiC/CCT chaperone network Jonathan J. Knowltona,b,1, Daniel Gestautc,1, Boxue Mad,e,f,2, Gwen Taylora,g,2, Alpay Burak Sevenh,i, Alexander Leitnerj, Gregory J. Wilsonk, Sreejesh Shankerl, Nathan A. Yatesm, B. V. Venkataram Prasadl, Ruedi Aebersoldj,n, Wah Chiud,e,f, Judith Frydmanc,3, and Terence S. Dermodya,g,o,3 aDepartment of Pediatrics, University of Pittsburgh School of Medicine, Pittsburgh, PA 15224; bDepartment of Pathology, Microbiology, and Immunology, Vanderbilt University Medical Center, Nashville, TN 37232; cDepartment of Biology, Stanford University, Stanford, CA 94305; dDepartment of Bioengineering, Stanford University, Stanford, CA 94305; eDepartment of Microbiology and Immunology, Stanford University, Stanford, CA 94305; fDepartment of Photon Science, Stanford University, Stanford, CA 94305; gCenter for Microbial Pathogenesis, UPMC Children’s Hospital of Pittsburgh, Pittsburgh, PA 15224; hDepartment of Structural Biology, Stanford University, Stanford, CA 94305; iDepartment of Molecular and Cellular Physiology, Stanford University, Palo Alto, CA 94305; jDepartment of Biology, Institute of Molecular Systems Biology, ETH Zürich, 8093 Zürich, Switzerland; kDepartment of Pediatrics, Vanderbilt University Medical Center, Nashville, TN 37232; lVerna and Marrs Mclean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX 77030; mDepartment of Cell Biology, University of Pittsburgh School of Medicine,
    [Show full text]
  • Gen-2020-0136.Pdf
    Genome Gut microbiome mediated epigenetic regulation of brain disorder and application of machine learning for multi- omics data analysis Journal: Genome Manuscript ID gen-2020-0136.R1 Manuscript Type: Mini Review Date Submitted by the 25-Sep-2020 Author: Complete List of Authors: Kaur, Harpreet; University of North Dakota School of Medicine and Health Sciences, Department of Biomedical Science Singh, Yuvraj; University of Calgary Faculty of Science, Department of Biological SciencesDraft Singh, Surjeet; University of Lethbridge, Department of Neuroscience, Canadian Centre for Behavioural Neuroscience (CCBN) Singh, Raja; University of Alberta, Faculty of Medicine and Dentistry; University of Calgary Cumming School of Medicine Gut-brain axis, epigenetics, neurodegenerative diseases, Machine Keyword: learning, Microbiota Is the invited manuscript for consideration in a Special Genome Biology Issue? : © The Author(s) or their Institution(s) Page 1 of 46 Genome 1 Gut microbiome mediated epigenetic regulation of brain disorder and application of 2 machine learning for multi-omics data analysis 3 4 Harpreet Kaur1*#, Yuvraj Singh2#, Surjeet Singh3, Raja B Singh4,5 5 1Department of Biomedical Sciences, School of Medicine and Health Sciences, University of 6 North Dakota, Grand Forks, ND, USA 7 2Department of Biological Sciences, Faculty of Science, University of Calgary, Alberta, Canada 8 3Department of Neuroscience, Canadian Centre for Behavioural Neuroscience (CCBN), 9 University of Lethbridge, Lethbridge, AB, Canada 10 4Faculty of Medicine & Dentistry, University of Alberta, Edmonton, Alberta, Canada 11 5Cumming School of Medicine, University of Calgary, Calgary, Alberta, Canada 12 13 # equal contribution 14 15 Draft 16 17 18 19 20 21 *Corresponding author: 22 Harpreet Kaur, PhD 23 Department of Biomedical Sciences 24 School of Medicine and Health Sciences 25 University of North Dakota 26 Grand Forks, ND, 58202, USA.
    [Show full text]
  • Maintenance of Mammary Epithelial Phenotype by Transcription Factor Runx1 Through Mitotic Gene Bookmarking Joshua Rose University of Vermont
    University of Vermont ScholarWorks @ UVM Graduate College Dissertations and Theses Dissertations and Theses 2019 Maintenance Of Mammary Epithelial Phenotype By Transcription Factor Runx1 Through Mitotic Gene Bookmarking Joshua Rose University of Vermont Follow this and additional works at: https://scholarworks.uvm.edu/graddis Part of the Biochemistry Commons, and the Genetics and Genomics Commons Recommended Citation Rose, Joshua, "Maintenance Of Mammary Epithelial Phenotype By Transcription Factor Runx1 Through Mitotic Gene Bookmarking" (2019). Graduate College Dissertations and Theses. 998. https://scholarworks.uvm.edu/graddis/998 This Thesis is brought to you for free and open access by the Dissertations and Theses at ScholarWorks @ UVM. It has been accepted for inclusion in Graduate College Dissertations and Theses by an authorized administrator of ScholarWorks @ UVM. For more information, please contact [email protected]. MAINTENANCE OF MAMMARY EPITHELIAL PHENOTYPE BY TRANSCRIPTION FACTOR RUNX1 THROUGH MITOTIC GENE BOOKMARKING A Thesis Presented by Joshua Rose to The Faculty of the Graduate College of The University of Vermont In Partial Fulfillment of the Requirements for the Degree of Master of Science Specializing in Cellular, Molecular, and Biomedical Sciences January, 2019 Defense Date: November 12, 2018 Thesis Examination Committee: Sayyed Kaleem Zaidi, Ph.D., Advisor Gary Stein, Ph.D., Advisor Seth Frietze, Ph.D., Chairperson Janet Stein, Ph.D. Jonathan Gordon, Ph.D. Cynthia J. Forehand, Ph.D. Dean of the Graduate College ABSTRACT Breast cancer arises from a series of acquired mutations that disrupt normal mammary epithelial homeostasis and create multi-potent cancer stem cells that can differentiate into clinically distinct breast cancer subtypes. Despite improved therapies and advances in early detection, breast cancer remains the leading diagnosed cancer in women.
    [Show full text]
  • A Concise Review of Human Brain Methylome During Aging and Neurodegenerative Diseases
    BMB Rep. 2019; 52(10): 577-588 BMB www.bmbreports.org Reports Invited Mini Review A concise review of human brain methylome during aging and neurodegenerative diseases Renuka Prasad G & Eek-hoon Jho* Department of Life Science, University of Seoul, Seoul 02504, Korea DNA methylation at CpG sites is an essential epigenetic mark position of carbon in the cytosine within CG dinucleotides that regulates gene expression during mammalian development with resultant formation of 5mC. The symmetrical CG and diseases. Methylome refers to the entire set of methylation dinucleotides are also called as CpG, due to the presence of modifications present in the whole genome. Over the last phosphodiester bond between cytosine and guanine. The several years, an increasing number of reports on brain DNA human genome contains short lengths of DNA (∼1,000 bp) in methylome reported the association between aberrant which CpG is commonly located (∼1 per 10 bp) in methylation and the abnormalities in the expression of critical unmethylated form and referred as CpG islands; they genes known to have critical roles during aging and neuro- commonly overlap with the transcription start sites (TSSs) of degenerative diseases. Consequently, the role of methylation genes. In human DNA, 5mC is present in approximately 1.5% in understanding neurodegenerative diseases has been under of the whole genome and CpG base pairs are 5-fold enriched focus. This review outlines the current knowledge of the human in CpG islands than other regions of the genome (3, 4). CpG brain DNA methylomes during aging and neurodegenerative islands have the following salient features. In the human diseases.
    [Show full text]
  • Architecture of the Human Mtorc2 Core Complex
    1 Architecture of the Human mTORC2 Core 2 Complex 3 4 5 6 7 8 Authors: 9 Edward Stuttfeld1*, Christopher H. S. Aylett2*, Stefan Imseng1, Daniel Boehringer2, Alain 10 Scaiola2, Evelyn Sauer1, Michael N. Hall1†, Timm Maier1†, Nenad Ban2† 11 12 Author affiliations: 13 1 Biozentrum, University of Basel, Klingelbergstrasse 50/70, 4056 Basel, Switzerland 14 2 Institute for Molecular Biology and Biophysics, ETH Zürich, Zürich, Switzerland 15 16 * These authors contributed equally to this work 17 18 † Correspondence should be addressed to M.N.H., T.M. or N.B. 19 E-mail: [email protected], [email protected] or [email protected] 20 21 22 1 23 Abstract 24 The mammalian target of rapamycin (mTOR) is a key protein kinase controlling cellular 25 metabolism and growth. It is part of the two structurally and functionally distinct multiprotein 26 complexes mTORC1 and mTORC2. Dysregulation of mTOR occurs in diabetes, cancer and 27 neurological disease. We report the architecture of human mTORC2 at intermediate 28 resolution, revealing a conserved binding site for accessory proteins on mTOR and 29 explaining the structural basis for the rapamycin insensitivity of the complex. 30 2 31 Introduction 32 The serine/threonine kinase mammalian target of rapamycin (mTOR) is the master 33 regulator of cellular growth (Saxton & Sabatini, 2017). Originally identified in yeast as TOR 34 (Heitman, Movva, & Hall, 1991; Kunz et al., 1993), the mammalian orthologue mTOR senses 35 a range of cellular cues, including nutrient and energy availability, and growth factor signals, 36 to control metabolism and autophagy (Laplante & Sabatini, 2012; Loewith & Hall, 2011; 37 Saxton & Sabatini, 2017).
    [Show full text]
  • The Role of the Chaperone CCT in Assembling Cell Signaling Complexes
    Brigham Young University BYU ScholarsArchive Theses and Dissertations 2020-07-21 The Role of the Chaperone CCT in Assembling Cell Signaling Complexes Nicole C. Tensmeyer Brigham Young University Follow this and additional works at: https://scholarsarchive.byu.edu/etd Part of the Physical Sciences and Mathematics Commons BYU ScholarsArchive Citation Tensmeyer, Nicole C., "The Role of the Chaperone CCT in Assembling Cell Signaling Complexes" (2020). Theses and Dissertations. 9192. https://scholarsarchive.byu.edu/etd/9192 This Dissertation is brought to you for free and open access by BYU ScholarsArchive. It has been accepted for inclusion in Theses and Dissertations by an authorized administrator of BYU ScholarsArchive. For more information, please contact [email protected]. The Role of the Molecular Chaperone CCT in Assembling Cell Signaling Complexes Nicole C. Tensmeyer A dissertation submitted to the faculty of Brigham Young University in partial fulfillment of the requirements for the degree of Doctor of Philosophy Barry M. Willardson, Chair Josh L. Andersen John C. Price Pam Van Ry Department of Chemistry and Biochemistry Brigham Young University Copyright © 2020 Nicole C. Tensmeyer All Rights Reserved ABSTRACT The Role of the Molecular Chaperone CCT in the Assembly of Signaling Complexes Nicole C. Tensmeyer Department of Chemistry and Biochemistry, BYU Doctor of Philosophy In order to function, proteins must be folded into their native shape. While this can sometimes occur spontaneously, the process can be hindered by thermodynamic barriers, trapped intermediates, and aggregation prone hydrophobic interactions. Molecular chaperones are proteins that help client proteins or substrates overcome these barriers so that they can be folded properly.
    [Show full text]
  • Identification of Macrophage Polarization-Related Genes As Biomarkers of Chronic Obstructive Pulmonary Disease Based on Bioinformatics Analyses
    Hindawi BioMed Research International Volume 2021, Article ID 9921012, 17 pages https://doi.org/10.1155/2021/9921012 Research Article Identification of Macrophage Polarization-Related Genes as Biomarkers of Chronic Obstructive Pulmonary Disease Based on Bioinformatics Analyses Yalin Zhao , Meihua Li, Yanxia Yang, Tao Wu, Qingyuan Huang, Qinghua Wu, and Chaofeng Ren Respiratory and Critical Care Medicine, Kunming First People’s Hospital, Kunming, Yunnan Province, China Correspondence should be addressed to Yalin Zhao; [email protected] and Chaofeng Ren; [email protected] Received 31 March 2021; Accepted 4 June 2021; Published 21 June 2021 Academic Editor: Nagarajan Raju Copyright © 2021 Yalin Zhao et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Objectives. Chronic obstructive pulmonary disease (COPD) is characterized by lung inflammation and remodeling. Macrophage polarization is associated with inflammation and tissue remodeling, as well as immunity. Therefore, this study attempts to investigate the diagnostic value and regulatory mechanism of macrophage polarization-related genes for COPD by bioinformatics analysis and to provide a new theoretical basis for experimental research. Methods. The raw gene expression profile dataset (GSE124180) was collected from the Gene Expression Omnibus (GEO) database. Next, a weighted gene coexpression network analysis (WGCNA) was conducted to screen macrophage polarization-related genes. The differentially expressed genes (DEGs) between the COPD and normal samples were generated using DESeq2 v3.11 and overlapped with the macrophage polarization-related genes. Moreover, functional annotations of overlapped genes were conducted by Database for Annotation, Visualization and Integrated Discovery (DAVID) Bioinformatics Resource.
    [Show full text]
  • Cryo-EM Structure of Human Mtor Complex 2
    www.nature.com/cr www.cell-research.com ARTICLE Cryo-EM structure of human mTOR complex 2 Xizi Chen1,2,3, Mengjie Liu1,2,3, Yuan Tian1,2,3, Jiabei Li1,2,3, Yilun Qi1,2,3, Dan Zhao1,2,3, Zihan Wu1,2,3, Min Huang4, Catherine C. L. Wong4,5,6, Hong-Wei Wang7, Jiawei Wang7, Huirong Yang1,2,3 and Yanhui Xu1,2,3,8 Mechanistic target of rapamycin (mTOR) complex 2 (mTORC2) plays an essential role in regulating cell proliferation through phosphorylating AGC protein kinase family members, including AKT, PKC and SGK1. The functional core complex consists of mTOR, mLST8, and two mTORC2-specific components, Rictor and mSin1. Here we investigated the intermolecular interactions within mTORC2 complex and determined its cryo-electron microscopy structure at 4.9 Å resolution. The structure reveals a hollow rhombohedral fold with a 2-fold symmetry. The dimerized mTOR serves as a scaffold for the complex assembly. The N-terminal half of Rictor is composed of helical repeat clusters and binds to mTOR through multiple contacts. mSin1 is located close to the FRB domain and catalytic cavity of mTOR. Rictor and mSin1 together generate steric hindrance to inhibit binding of FKBP12-rapamycin to mTOR, revealing the mechanism for rapamycin insensitivity of mTORC2. The mTOR dimer in mTORC2 shows more compact conformation than that of mTORC1 (rapamycin sensitive), which might result from the interaction between mTOR and Rictor-mSin1. Structural comparison shows that binding of Rictor and Raptor (mTORC1-specific component) to mTOR is mutually exclusive. Our study provides a basis for understanding the assembly of mTORC2 and a framework to further characterize the regulatory mechanism of mTORC2 pathway.
    [Show full text]