DOCSLIB.ORG

Structural Insights Into Protein Arginine Symmetric Dimethylation by PRMT5

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Structural Insights Into Protein Arginine Symmetric Dimethylation by PRMT5 Structural insights into protein arginine symmetric dimethylation by PRMT5 Litao Suna,b,1,MingzhuWanga,1, Zongyang Lva,b, Na Yanga,YingfangLiua, Shilai Baoc,WeiminGonga,2, and Rui-Ming Xua,2 aNational Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China; bGraduate University of Chinese Academy of Sciences, Beijing 100049, China; and cState Key Laboratory of Molecular and Developmental Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China Edited by* Dinshaw J. Patel, Memorial Sloan-Kettering Cancer Center, New York, NY, and approved October 19, 2011 (received for review May 2, 2011) Symmetric and asymmetric dimethylation of arginine are isomeric enzyme shares high sequence homology with its human counter- protein posttranslational modifications with distinct biological part, and the recombinant protein displays robust and specific effects, evidenced by the methylation of arginine 3 of histone H4 symmetric arginine dimethylase activity in vitro (Fig. 1). The (H4R3): symmetric dimethylation of H4R3 leads to repression of structure shows the PRMT5 is composed of four clearly defined gene expression, while asymmetric dimethylation of H4R3 is asso- domains, a previously unsuspected TIM-barrel at the N-terminal ciated with gene activation. The enzymes catalyzing these modifi- end, a middle Rossmann-fold domain, a C-terminal β-barrel cations share identifiable sequence similarities, but the relationship domain, and a ∼60 residue dimerization domain inserted be- between their catalytic mechanisms is unknown. Here we analyzed tween β1 and β2 of the β-barrel domain (Fig. 2A). The first three the structure of a prototypic symmetric arginine dimethylase, domains are packed in a triangular manner, with direct contacts PRMT5, and discovered that a conserved phenylalanine in the between sequential domains, and the oligomerization domain active site is critical for specifying symmetric addition of methyl bridges the TIM-barrel and β-barrel domains. The structure of groups. Changing it to a methionine significantly elevates the the SAH-bound PRMT5 differs from that of free protein in overall methylase activity, but also converts PRMT5 to an enzyme that a N-terminal loop (L0) and helix (αA) are ordered in the that catalyzes both symmetric and asymmetric dimethylation of SAH-bound structure. We will use the SAH-bound structure for arginine. Our results demonstrate a common catalytic mechanism analysis unless explicitly noted. BIOCHEMISTRY intrinsic to both symmetric and asymmetric arginine dimethylases, PRMT5 exists as a homodimer, shown both in the crystal struc- and show that steric constrains in the active sites play an essential ture and in solution, as determined by analytic ultracentrifugation role in determining the product specificity of arginine methylases. (Fig. 2B, Fig. S1). The dimeric interface buries a total pair wise 2 This discovery also implies a potentially regulatable outcome of surface area of 2;305 Å , and the intermolecular interactions arginine dimethylation that may provide versatile control of eukar- occur between the dimerization domains and that between the yotic gene expression. TIM-barrel and β-barrel domains. Human PRMT5 has a shorter oligomerization domain, but extensive conservation of amino histone methylation ∣ transtriptional regulation ∣ RNA splicing ∣ acids involved in intermolecular interactions implies that it also crystal structure forms a dimer, consistent with the report of dimeric and higher oligomeric forms of human PRMT5 (5). In fact, all arginine methy- rotein arginine methyltransferase 5 (PRMT5) catalyzes the lases with known structures dimerize via a homologous dimeriza- Pevenly addition of two methyl groups to the two ω-guanidino tion domain, also known as the dimerization “arm” (Fig. 3A), G 0G nitrogen atoms of arginine, resulting in ω-N , N symmetric (22–26). Thus, protein dimerization appears to be an evolutionarily dimethylation of arginine (sDMA) of the target protein (1–5). conserved property of arginine methylases, although the functional PRMT5 functions in the nucleus as well as in the cytoplasm, significance remains poorly understood. and its substrates include histones, spliceosomal proteins, tran- scription factors, and proteins involved in piRNA biogenesis Comparison with Arginine Asymmetric Dimethylases. The overall (6). Symmetric dimethylation of these proteins profoundly impact fold and spatial positioning of the Rossmann-fold and β-barrel many biological processes; e.g., epigenetic control of gene expres- domains are similar to that of type-I enzymes, represented by sion (7), splicing regulation (2, 3, 8, 9), circadian rhythms (9, 10), PRMT1 and CARM1 (Fig. 3A) (24–26). The root-mean-squared DNA damage response (11, 12), and germ cell development and deviation of Cα positions between PRMT5 and PRMT1 is pluripotency (13–16). Interestingly, both PRMT5 and a group of approximately 2.1 Å when both the Rossmann-fold and β-barrel asymmetric (type-I) arginine dimethylases, which add two methyl domains are compared, whereas it is 1.4 Å for the Rossmann-fold groups to the same ω-guanidino nitrogen atom (aDMA), share domain alone. In particular, a segment including a N-terminal common recognition sequences, and the target arginine can often loop (L0) and a following helix (αA) (a.a. 359–380) became be symmetrically or asymmetrically dimethylated. Yet, these iso- ordered upon SAH binding. Helix αA is positioned similar to that meric modifications have distinct biological effects. One such ex- ample occurs at arginine-3 of histone H4 (H4R3). Symmetric dimethylation of H4R3 has been linked to repression of gene Author contributions: L.S., Y.L., S.B., W.G., and R.-M.X. designed research; L.S., M.W., Z.L., – N.Y., and R.-M.X. performed research; S.B. contributed new reagents/analytic tools; L.S., expression (17 19), while asymmetric dimethylation of H4R3 is M.W., Z.L., N.Y., Y.L., S.B., W.G., and R.-M.X. analyzed data; and L.S., M.W., and R.-M.X. associated with gene activation (20, 21). The startling difference wrote the paper. in biological effects of sDMA and aDMA modifications necessi- The authors declare no conflict of interest. tates the understanding of the enzymatic mechanisms differentiat- *This Direct Submission article had a prearranged editor. ing the two chemically isomeric but functionally antagonistic Data deposition: The atomic coordinates have been deposited in the Protein Data Bank, posttranslational modifications. www.pdb.org (PDB ID codes 3UA3 and 3UA4). 1L.S. and M.W. contributed equally to this work. Results 2To whom correspondence may be addressed. E-mail: [email protected] or wgong@ Overall Structure. We have determined the crystal structures of sun5.ibp.ac.cn. full-length PRMT5 from Caenorhabditis elegans, alone and in This article contains supporting information online at www.pnas.org/lookup/suppl/ complex with S-Adenosyl-L-homocysteine (SAH). The nematode doi:10.1073/pnas.1106946108/-/DCSupplemental. www.pnas.org/cgi/doi/10.1073/pnas.1106946108 PNAS Early Edition ∣ 1of6 Downloaded by guest on September 29, 2021 Fig. 1. Structural and functional conservation of PRMT5. (A) A schematic representation of domain structures of C. elegans and human PRMT5, and a re- presentative type-I arginine methylase, PRMT1 of rat. The lengths of the boxes are approximately drawn in scale with the protein lengths, and the residue numbers at domain boundaries are labeled. Areas filled in tan, cyan, green, and yellow represent TIM-barrel, Rossmann-fold, β-barrel, and oligomerization domains, respectively. Levels of amino acid identity and similarity of the individual domains between C. elegans and human PRMT5s, and that between human PRMT5 and rat PRMT1 are shown. (B) Sequence alignment. The full-length sequences of C. elegans and human PRMT5s, and the regions of the solved structures of rat PRMT1 and mouse CARM1 are aligned. Residues conserved in all four proteins are shown in white letters over purple background, and similar residues are indicated with red letters. Residues conserved in PRMT5 proteins and type-I arginine methylases are highlighted tan and yellow, respectively. Blue stars mark the CePRMT5 residues subjected to mutagenesis. At the top of the sequences, a schematic representation of the secondary structure elements of CePRMT5 is shown. Every ten residues are indicated with a “·” sign. (C) Enzymatic activity assay. Top box, coomassie-stained gel of enzymes and substrate (histone H4) used. GST-tagged rat PRMT1 and poly(His)-tagged C. elegans PRMT5 were expressed in E. coli, and flag-tagged human PRMT5 was purified from HEK293 cells. Approximately 5 μg of enzymes and histone H4 each were used in the assay. Top 2nd box, autoradiograph generated with the use of 0.25 mCi of SAM with tritiated methyl group. Top 3rd box, Western blot detection of asymmetrically dimethylated histone H4R3. Bottom box, Western blot detection of symme- trically dimethylated histone H4R3. found in type-I enzymes, sheltering SAH from exposing to the PRMT5 proteins (Pro366, Leu367, and Leu371) and forms a solvent and creating a secluded catalytic active site. However, solvent inaccessible area for catalysis; (ii) the N-terminal end key residues responsible for the disordered-to-ordered conforma- of L0 makes a U-turn and contacts the dimerization domain, tional transition upon SAH/SAM binding are separately con- which stabilizes the loop in a conformation endowed with the served among PRMT5 family members (Fig. 1B). Among which, ability to influence substrate binding (Figs. 3 A and B). Hence, Tyr376 and Phe379 on αA interact with the ribose and homocys- the highly conserved PRMT5 loop is important for SAH/SAM teine moieties via hydrogen bonds and van der Waals contacts, binding, as well as in a position to regulate substrate binding. respectively. Loop L0 contains several amino acids uniquely conserved in PRMT5s across species. The corresponding region The Active Site. The active site of PRMT5 is identified by the loca- in PRMT1 is disordered, and that of CARM1 adopts a helical tion of the sulfur atom of SAH and a pair of invariant glutamate conformation.
Load more

Recommended publications
  • Gene Symbol Gene Description ACVR1B Activin a Receptor, Type IB
    Table S1. Kinase clones included in human kinase cDNA library for yeast two-hybrid screening Gene Symbol Gene Description ACVR1B activin A receptor, type IB ADCK2 aarF domain containing kinase 2 ADCK4 aarF domain containing kinase 4 AGK multiple substrate lipid kinase;MULK AK1 adenylate kinase 1 AK3 adenylate kinase 3 like 1 AK3L1 adenylate kinase 3 ALDH18A1 aldehyde dehydrogenase 18 family, member A1;ALDH18A1 ALK anaplastic lymphoma kinase (Ki-1) ALPK1 alpha-kinase 1 ALPK2 alpha-kinase 2 AMHR2 anti-Mullerian hormone receptor, type II ARAF v-raf murine sarcoma 3611 viral oncogene homolog 1 ARSG arylsulfatase G;ARSG AURKB aurora kinase B AURKC aurora kinase C BCKDK branched chain alpha-ketoacid dehydrogenase kinase BMPR1A bone morphogenetic protein receptor, type IA BMPR2 bone morphogenetic protein receptor, type II (serine/threonine kinase) BRAF v-raf murine sarcoma viral oncogene homolog B1 BRD3 bromodomain containing 3 BRD4 bromodomain containing 4 BTK Bruton agammaglobulinemia tyrosine kinase BUB1 BUB1 budding uninhibited by benzimidazoles 1 homolog (yeast) BUB1B BUB1 budding uninhibited by benzimidazoles 1 homolog beta (yeast) C9orf98 chromosome 9 open reading frame 98;C9orf98 CABC1 chaperone, ABC1 activity of bc1 complex like (S. pombe) CALM1 calmodulin 1 (phosphorylase kinase, delta) CALM2 calmodulin 2 (phosphorylase kinase, delta) CALM3 calmodulin 3 (phosphorylase kinase, delta) CAMK1 calcium/calmodulin-dependent protein kinase I CAMK2A calcium/calmodulin-dependent protein kinase (CaM kinase) II alpha CAMK2B calcium/calmodulin-dependent
    [Show full text]
  • Riok1 (BC002158) Mouse Tagged ORF Clone – MR204785 | Origene
    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 MR204785 Riok1 (BC002158) Mouse Tagged ORF Clone Product data: Product Type: Expression Plasmids Product Name: Riok1 (BC002158) Mouse Tagged ORF Clone Tag: Myc-DDK Symbol: Riok1 Synonyms: 3110046C13Rik, Ad034, MGC7300 Vector: pCMV6-Entry (PS100001) E. coli Selection: Kanamycin (25 ug/mL) Cell Selection: Neomycin ORF Nucleotide >MR204785 ORF sequence Sequence: Red=Cloning site Blue=ORF Green=Tags(s) TTTTGTAATACGACTCACTATAGGGCGGCCGGGAATTCGTCGACTGGATCCGGTACCGAGGAGATCTGCC GCCGCGATCGCC ATGGTGAGGACGTGGGCAGAGAAGGAGATGAGGAATTTGTGCAGGCTAAAAACAGCAAACATACCATGTC CAGAACCAATCAGGCTAAGAAGTCATGTTCTTCTCATGGGCTTCATTGGCAAGGATGACATGCCAGCCCC ACTTTTGAAAAATGTCCAGCTGTCAGAGTCCAAGGCACGGGAGTTGTACCTGCAGGTCATTCAGTACATG AGGAAAATGTATCAGGATGCTAGACTTGTCCACGCGGATCTCAGTGAATTCAACATGCTGTACCATGGTG GAGATGTTTACATCATTGATGTTTCTCAGTCTGTGGAGCATGACCACCCACATGCATTGGAGTTCTTGAG AAAAGACTGTACCAATGTCAATGATTTCTTTTCCAAGCATGCTGTTGCAGTGATGACCGTGCGGGAGCTC TTCGACTTCGTCACAGATCCCTCCATCACTGCTGACAACATGGATGCTTACCTGGAAAAGGCTATGGAAA TAGCATCCCAGAGGACCAAGGAAGAAAAGACTAGCCAAGATCATGTGGATGAAGAGGTGTTCAAACAAGC ATATATTCCCAGAACCTTAAACGAAGTAAAGAATTATGAGAGAGATGTGGACATCATGATGAGGTTAAAG GAAGAAGACATGGCTTTGAACACTCAGCAAGACAACATTCTATACCAGACTGTCATGGGATTGAAAAAAG ATTTGTCAGGAGTCCAGAAGGTCCCCGCGCTCCTAGAAAGTGAAGTTAAGGAAGAGACTTGTTTTGGTTC AGACGATGCTGGGGGCTCTGAGTGCTCCGACACAGTCTCTGAAGAGCAGGAAGATCAAGCCGGATGCAGA AACCATATTGCTGACCCCGACGTTGATAAAAAGGAAAGAAAAAAGATGGTCAAGGAAGCCCAGAGAGAGA
    [Show full text]
  • Human RIOK1 / RIO Kinase 1 Protein (His & GST Tag)
    Human RIOK1 / RIO kinase 1 Protein (His & GST Tag) Catalog Number: 14477-H20B General Information SDS-PAGE: Gene Name Synonym: AD034; bA288G3.1; RRP10; 3110046C13Rik; 5430416A05Rik; Ad034 Protein Construction: A DNA sequence encoding the human RIOK1 (Q9BRS2) (Met1-Lys568) was expressed with the N-terminal polyhistidine-tagged GST tag at the N-terminus. Source: Human Expression Host: Baculovirus-Insect Cells QC Testing Purity: > 85 % as determined by SDS-PAGE Bio Activity： Protein Description Kinase activity untested RIOK1, also known as RIO kinase 1, is a member of the RIO family of Endotoxin: atypical serine protein kinases first characterized in yeast. RIOK1 and RIOK2 proteins are present in organisms from Archaea to humans. RIOK1 < 1.0 EU per μg of the protein as determined by the LAL method functios as a new interactor of protein arginine methyltransferase 5 (PRMT5), competes with pICln for binding and modulates PRMT5 complex Stability: composition and substrate specificity. RioK1 and pICln bind to PRMT5 in a Samples are stable for up to twelve months from date of receipt at -70 ℃ mutually exclusive fashion. This results in a PRMT5-WD45/MEP50 core structure that either associates with pICln or RioK1 RIOK1 in distinct Predicted N terminal: Met complexes. RIOK1 functions in analogy to pICln as an adapter protein by recruiting the RNA-binding protein nucleolin to the PRMT5 complex for its Molecular Mass: symmetrical methylation. The recombinant human RIOK1/GST chimera consists of 805 amino acids References and has a calculated molecular mass of 93.4 kDa. The recombinant protein migrates as an approximately 94 kDa band in SDS-PAGE under reducing 1.Widmann B.
    [Show full text]
  • Biochemical Investigation of the Interaction of Picln, Riok1 and COPR5 with the PRMT5–MEP50 Complex
    Communications ChemBioChem doi.org/10.1002/cbic.202100079 1 2 3 Biochemical Investigation of the Interaction of pICln, RioK1 4 5 and COPR5 with the PRMT5–MEP50 Complex 6 [a, b] [c] [d, e] ' [f] 7 Adrian Krzyzanowski, Raphael Gasper, Hélène Adihou, Peter t Hart,* and [a, b] 8 Herbert Waldmann* 9 10 compete for the same binding site, but are not competitive 11 The PRMT5–MEP50 methyltransferase complex plays a key role with MEP50.[5] 12 in various cancers and is regulated by different protein–protein PRMT5 contains three domains, that is, the catalytic site- 13 interactions. Several proteins have been reported to act as containing Rossmann fold, a β-barrel domain used for dimeriza- 14 adaptor proteins that recruit substrate proteins to the active tion, and the TIM barrel, which acts as interaction site for 15 site of PRMT5 for the methylation of arginine residues. To MEP50 (Figure 1A).[3a,c] PRMT5 and MEP50 form a hetero- 16 define the interaction between these adaptor proteins and octameric complex whose structure has been determined,[3a] 17 PRMT5, we employed peptide truncation and mutation studies and structural information on the interaction between PRMT5 18 and prepared truncated protein constructs. We report the and the other adaptor proteins has only very recently been 19 characterisation of the interface between the TIM barrel of described in preliminary form.[7] Overexpression of PRMT5 is 20 PRMT5 and the adaptor proteins pICln, RioK1 and COPR5, and frequently observed in cancer which makes the protein an 21 identify the consensus amino acid sequence GQF[D/E]DA[E/D] intensively investigated anticancer target, and active-site di- 22 involved in binding.
    [Show full text]
  • Targeting Posttranslational Modifications of RIOK1
    RESEARCH ARTICLE Targeting posttranslational modifications of RIOK1 inhibits the progression of colorectal and gastric cancers Xuehui Hong1,2,3,4†, He Huang5,6†, Xingfeng Qiu2,3,4, Zhijie Ding2,3,4, Xing Feng7, Yuekun Zhu8, Huiqin Zhuo2,3,4, Jingjing Hou2,3,4, Jiabao Zhao2,3,4, Wangyu Cai2,3,4, Ruihua Sha9, Xinya Hong10, Yongxiang Li11*, Hongjiang Song12*, Zhiyong Zhang1,13* 1Longju Medical Research Center, Key Laboratory of Basic Pharmacology, Ministry of Education, Zunyi Medical College, Zunyi, China; 2Department of Gastrointestinal Surgery, Zhongshan Hospital of Xiamen University, Xiamen, China; 3Institute of Gastrointestinal Oncology, Medical College of Xiamen University, Xiamen, China; 4Xiamen Municipal Key Laboratory of Gastrointestinal Oncology, Xiamen, China; 5Department of Histology and Embryology, Xiangya School of Medicine, Central South University, Changsha, China; 6Digestive Cancer Laboratory, Second Affiliated Hospital of Xinjiang Medical University, Urumqi, China; 7Department of Radiation Oncology, Cancer Institute of New Jersey, Rutgers University, New Brunswick, United States; 8Department of General Surgery, The First Affiliated Hospital of Harbin Medical University, Harbin, China; 9Department of Digestive Disease, Hongqi Hospital, Mudanjiang Medical University, Mudanjiang, China; 10Department *For correspondence: of Medical Imaging and Ultrasound, Zhongshan Hospital of Xiamen University, 11 [email protected] Xiamen, Fujian, China; Department of General Surgery, The First Affiliated (YL); Hospital of Anhui Medical University, Hefei, China; 12Department of General [email protected] Surgery, The Third Affiliated Hospital of Harbin Medical University, Harbin, China; (HS); 13Department of Surgery, Robert-Wood-Johnson Medical School University [email protected] (ZZ) † Hospital, Rutgers University, The State University of New Jersey, New Brunswick, These authors contributed United States equally to this work Competing interests: The authors declare that no competing interests exist.
    [Show full text]
  • Genomic Alterations Link Rho Family of Gtpases to the Highly Invasive Phenotype of Pancreas Cancer
    Genomic alterations link Rho family of GTPases to the highly invasive phenotype of pancreas cancer Alec C. Kimmelmana,b, Aram F. Hezela,c, Andrew J. Aguirrea, Hongwu Zhenga, Ji-hye Paika, Haoqiang Yinga, Gerald C. Chua, Jean X. Zhanga,d, Ergun Sahina, Giminna Yeod, Aditya Ponugotid, Roustem Nabioullind, Scott Derooa, Shenghong Yangb, Xiaoxu Wangb, John P. McGrathd, Marina Protopopovad, Elena Ivanovad, Jianhua Zhangd, Bin Fengd, Ming S. Tsaoe, Mark Redstonf, Alexei Protopopovd, Yonghong Xiaod, P. Andrew Futrealg, William C. Hahna,h,i, David S. Klimstraj, Lynda China,d,k, and Ronald A. DePinhoa,d,h,1 aDepartment of Medical Oncology and bHarvard Radiation Oncology Program, Dana-Farber Cancer Institute and Harvard Medical School, Boston, MA 02115; dCenter for Applied Cancer Science of the Belfer Institute for Innovative Cancer Science, Dana-Farber Cancer Institute, Boston, MA 02115; kDepartment of Dermatology, Brigham and Women’s Hospital and Harvard Medical School, Boston, MA 02115; hDepartments of Medicine and Genetics, Harvard Medical School, Boston, MA 02115; eUniversity Health Network, Ontario Cancer Institute and Princess Margaret Hospital, Toronto, ON, Canada M5G 2M9; jDepartment of Pathology, Memorial Sloan Kettering Cancer Center, New York, NY 10065; fMolecular Diagnostics, Ameripath, Newton, MA 02464; gCancer Genome Project, Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge CB10 1SA, United Kingdom; iBroad Institute of Harvard and MIT, Cambridge, MA 02142; and cTucker Gosnell Center for Gastrointestinal Cancers, Massachusetts General Hospital Cancer Center, Lawrence House, P.O. Box 232, Boston, MA 02114 Communicated by David M. Livingston, Dana–Farber Harvard Cancer Center, Boston, MA, October 10, 2008 (received for review May 7, 2008) Pancreas ductal adenocarcinoma (PDAC) is a highly lethal cancer that high-resolution aCGH to better define the atlas of recurrent CNAs typically presents as advanced, unresectable disease.
    [Show full text]
  • Co-Overexpression of RIOK1 and AKT1 As a Prognostic Risk Factor In
    Journal of Cancer 2021, Vol. 12 5745 Ivyspring International Publisher Journal of Cancer 2021; 12(19): 5745-5752. doi: 10.7150/jca.60596 Research Paper Co-overexpression of RIOK1 and AKT1 as a prognostic risk factor in glioma Yiwei Wang1, 2, Xiaochen Xie3, Shu Li1, Dongyong Zhang4, Heyu Zheng1, Min Zhang2, Zhong Zhang2 1. Department of Human Anatomy, Shenyang Medical College, Shenyang City, Liaoning Province 110034, P.R. China. 2. Department of Pathology, College of Basic Medical Sciences, Shenyang Medical College, Shenyang City, Liaoning Province 110034, P.R. China. 3. Department of Endocrinology and Metabolism, Institute of Endocrinology, Liaoning Provincial Key Laboratory of Endocrine Diseases, The First Affiliated Hospital of China Medical University, China Medical University, Shenyang, Liaoning, 110001, P.R. China. 4. Department of Neurosurgery, First Affiliated Hospital of China Medical University, Heping District, Shenyang City, Liaoning Province, 110001, P.R. China Corresponding author: E-mail: [email protected] © The author(s). This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/). See http://ivyspring.com/terms for full terms and conditions. Received: 2021.03.18; Accepted: 2021.07.19; Published: 2021.07.25 Abstract Glioblastoma multiforme (GBM) is one of the most frequent primary malignancies of the brain. Although the treatment strategy has significantly improved, patient prognosis remains poor. In vitro studies have shown that the right open reading frame kinase 1/protein kinase B (RIOK1-AKT) signaling pathway plays an important role in the malignant phenotype of glioma cells. This study aimed to investigate the co-expression of RIOK1 and ATK in glioma tissues and its clinical significance.
    [Show full text]
  • RIOK1 Kinase Activity Is Required for Cell Survival Irrespective of MTAP Status
    www.oncotarget.com Oncotarget, 2018, Vol. 9, (No. 47), pp: 28625-28637 Research Paper RIOK1 kinase activity is required for cell survival irrespective of MTAP status Alexandra Hörmann1,*, Barbara Hopfgartner1,*, Thomas Köcher2, Maja Corcokovic1, Teresa Krammer1, Christoph Reiser1, Gerd Bader1, Junwei Shi3, Katharina Ehrenhöfer1, Simon Wöhrle1, Norbert Schweifer1, Christopher R. Vakoc3, Norbert Kraut1, Mark Pearson1, Mark Petronczki1 and Ralph A. Neumüller1 1Boehringer Ingelheim RCV GmbH & Co KG, 1120 Vienna, Austria 2Vienna Biocenter Core Facilities GmbH, 1030 Vienna, Austria 3Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724, USA *These authors have contributed equally to this work Correspondence to: Ralph A. Neumüller, email: [email protected] Keywords: RIOK1; MTAP; PRMT5; cancer; target Received: November 16, 2017 Accepted: May 19, 2018 Published: June 19, 2018 Copyright: Hörmann et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License 3.0 (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. ABSTRACT Genotype specific vulnerabilities of cancer cells constitute a promising strategy for the development of new therapeutics. Deletions of non-essential genes in tumors can generate unique vulnerabilities which could be exploited therapeutically. The MTAP gene is recurrently deleted in human cancers because of its chromosomal proximity to the tumor suppressor gene CDKN2A. Recent studies have uncovered an increased dependency of MTAP-deleted cancer cells on the function of a PRMT5 containing complex, including WDR77, PRMT5 and the kinase RIOK1. As RIOK1 kinase activity constitutes a potential therapeutic target, we wanted to test if MTAP deletion confers increased sensitivity to RIOK1 inhibition.
    [Show full text]
  • RIOK2 and Other Trans-​Acting Factors (Nat
    NEWS & ANALYSIS BIOBUSINESS BRIEFS TARGET WATCH release of RIOK2 and other trans- acting factors (Nat. Struct. Mol. Biol. 19, 1316–1323; 2012). RIOK2 has also been linked to AKT RIOK2: straddling the kinase/ATPase line signalling in glioblastoma (PLoS Genet. 9, e1003253; 2013). RIOK2 and RIOK1 The RIO kinases (RIOKs) are a family Biological functions were found in a complex with TORC2, of atypical kinases present in eukaryotes RIOK2 is essential in yeast, mice and in and overexpression of RIOK2 resulted in and archaea (Genome Res. 8, 1038–1047; at least 560 human cell lines (see DepMap increased phosphorylation of AKT S473, 1998). RIOK1 and RIOK2 are conserved in Related links). Essentiality is largely a TORC2 substrate. In turn, activation of throughout evolution, whereas multicellular attributed to the role of RIOK2 in small AKT resulted in stabilization of RIOK2 eukaryotes, including humans, have a third ribosomal subunit maturation, a complex protein levels creating a feed-forward RIOK, RIOK3. All three RIOKs are found process involving more than 150 trans- activation loop. Although untested, RIOK2 in the pre-40S ribosome, and RIOK1 and acting factors in the nucleus and cytoplasm. has a putative AKT consensus sequence with RIOK2 are essential for ribosomal maturation RIOK2 functions as a trans- acting factor documented phosphorylation. (Mol. Cell. Biol. 23, 2083–2095; 2003), that shepherds the last steps of maturation but their functions are non-overlapping and of the 40S subunit. Depletion of RIOK2 Chemical tools poorly understood mechanistically. More results in a late pre-40S-biogenesis defect. Broad kinase screening has demonstrated that recently, RIOKs have been found to regulate In the nucleus, RIOK2 binds the pre-40S RIOK2 is competent to bind small molecules other biological pathways including cell cycle subunit and helps facilitate export through (PLoS One 12, e0181585; 2017).
    [Show full text]
  • 95742748.Pdf
    EBioMedicine 20 (2017) 79–97 Contents lists available at ScienceDirect EBioMedicine journal homepage: www.ebiomedicine.com Research Paper The Atypical Kinase RIOK1 Promotes Tumor Growth and Invasive Behavior Florian Weinberg a,b,c,NadineReischmanna,b,d,LisaFauthe,#, Sanaz Taromi f,#, Justin Mastroianni b,f,#, Martin Köhler a,b,d, Sebastian Halbach a,b,d, Andrea C. Becker g,h, Niantao Deng i,j, Tatjana Schmitz a, Franziska Maria Uhl a,b,f, Nicola Herbener e, Bianca Riedel e, Fabian Beier e, Alexander Swarbrick i,j, Silke Lassmann c,e,k, Jörn Dengjel c,g,h,l, Robert Zeiser c,f, Tilman Brummer a,b,c,k,⁎ a Institute of Molecular Medicine and Cell Research (IMMZ), Faculty of Medicine, Albert-Ludwigs-University (ALU), Freiburg, Germany b Faculty of Biology, ALU, Freiburg, Germany c BIOSS Centre for Biological Signalling Studies, BIOSS, ALU, Germany d Spemann Graduate School of Biology and Medicine (SGBM), ALU, Freiburg, Germany e Institute for Surgical Pathology, Medical Center and Faculty of Medicine, ALU, Germany f Department of Hematology and Oncology, University Medical Center, ALU, Freiburg, Germany g Freiburg Institute for Advanced Studies (FRIAS), ALU, Freiburg, Germany h Department of Dermatology, University Medical Center - ALU, Freiburg, Germany i Garvan Institute of Medical Research, Darlinghurst, New South Wales, Australia j St Vincent's Clinical School, Faculty of Medicine, UNSW, Sydney, Australia k German Cancer Consortium (DKTK, Freiburg) and German Cancer Research Center (DKFZ), Heidelberg, Germany l Department of Biology, University of Fribourg, Fribourg, Switzerland article info abstract Article history: Despite being overexpressed in different tumor entities, RIO kinases are hardly characterized in mammalian cells.
    [Show full text]
  • IEEE Conference Paper Template
    Two-Column Template for LACCEI Conference Proceedings Based on IEEE Format (Title) Albert Djoum1, Hongpeng Wang1, and Nicole Laronde, PhD1 1University of Maryland, College Park, United States, [email protected] ,[email protected], [email protected] Abstract DIFFERENCE BETWEEN A NORMAL OVARY AND ONE AFFECTED For all cancers, increased ribosome biogenesis is a critical BY CANCER.2 requirement and RIO kinases play a critical role in maintaining processing efficiency. RioK1 is important because it is required for cytoplasmic maturation of the small(40S) subunit. RioK1 is also important to other cancer promoting processes, such as proliferation and migration. In functions other than ribosome biogenesis, RioK1 is used as an adaptor protein of PRMT5 (protein arginine methyltransferase 5) and recruits nucleolin for symmetrical demethylation. In relation to cancer, however, specific roles, as well as structure for the RioK1-PRMT5-nucleolin complex is still unknown. Overall, this research will investigate the effects of RIOK1 knockdown and overexpression on cancer-supporting processes such as proliferation, migration, and invasion. The goal of this specific project was as such: what is the ideal molar ratio of mixing RioK-1 and PRMT5, that results in the best, most homogeneous, complex being formed? After determining this ratio, X-ray crystallography will be used on the RioK1-PRMT5-nucleolin complex and the structure will be determined. The methodology used was as follows: first, Riok1 and PRMT5 from Chaetomium Thermophilum were expressed in E. coli bacteria and then purified. Afterwards, RioK1 and PRMT5 were mixed in various molar ratios and then analyzed on native PAGE gel. Initially, RioK1 and PRMT5 were both varied, but then RioK1 was kept at a constant FIGURE 1.
    [Show full text]
  • Precision Medicine for Hepatocelluar Carcinoma Using Molecular Pattern Diagnostics: Results from a Preclinical Pilot Study
    Citation: Cell Death and Disease (2017) 8, e2867; doi:10.1038/cddis.2017.229 OPEN Official journal of the Cell Death Differentiation Association www.nature.com/cddis Precision medicine for hepatocelluar carcinoma using molecular pattern diagnostics: results from a preclinical pilot study Rahul Agarwal1, Yuan Cao2, Klaus Hoffmeier1, Nicolas Krezdorn1, Lukas Jost1, Alejandro Rodriguez Meisel1, Ruth Jüngling1, Francesco Dituri2, Serena Mancarella2, Björn Rotter1, Peter Winter1 and Gianluigi Giannelli*,2 The aim of this study was to design a road map for personalizing cancer therapy in hepatocellular carcinoma (HCC) by using molecular pattern diagnostics. As an exploratory study, we investigated molecular patterns of tissues of two tumors from individual HCC patients, which in previous experiments had shown contrasting reactions to the phase 2 transforming growth factor beta receptor 1 inhibitor galunisertib. Cancer-driving molecular patterns encompass – inter alias – altered transcription profiles and somatic mutations in coding regions differentiating tumors from their respective peritumoral tissues and from each other. Massive analysis of cDNA ends and all-exome sequencing demonstrate a highly divergent transcriptional and mutational landscape, respectively, for the two tumors, that offers potential explanations for the tumors contrasting responses to galunisertib. Molecular pattern diagnostics (MPDs) suggest alternative, individual-tumor-specific therapies, which in both cases deviate from the standard sorafenib treatment and from each other. Suggested personalized therapies use kinase inhibitors and immune- focused drugs as well as low-toxicity natural compounds identified using an advanced bioinformatics routine included in the MPD protocol. The MPD pipeline we describe here for the prediction of suitable drugs for treatment of two contrasting HCCs may serve as a blueprint for the design of therapies for various types of cancer.
    [Show full text]