title: NEDDylation promotes stress granule assembly.
abstract: Stress granules (SGs) harbour translationally stalled messenger ribonucleoproteins and play important roles in regulating gene expression and cell fate. Here we show that neddylation promotes SG assembly in response to arsenite-induced oxidative stress. Inhibition or depletion of key components of the neddylation machinery concomitantly inhibits stress-induced polysome disassembly and SG assembly. Affinity purification and subsequent mass-spectrometric analysis of Nedd8-conjugated proteins from translationally stalled ribosomal fractions identified ribosomal proteins, translation factors and RNA-binding proteins (RBPs), including SRSF3, a previously known SG regulator. We show that SRSF3 is selectively neddylated at Lys85 in response to arsenite. A non-neddylatable SRSF3 (K85R) mutant do not prevent arsenite-induced polysome disassembly, but fails to support the SG assembly, suggesting that the neddylation pathway plays an important role in SG assembly.
authors.affiliation: Department of Cellular &Molecular Medicine, College of Medicine, Chosun University, Gwangju 61452, Republic of Korea.
authors.forename: Aravinth Kumar
authors.initials: AK
authors.lastname: Jayabalan
authors.affiliation: Department of Cell Biology, Microbiology, and Molecular Biology, College of Arts and Sciences, University of South Florida, Tampa, Florida 33620, USA.
authors.forename: Anthony
authors.initials: A
authors.lastname: Sanchez
authors.affiliation: Department of Cellular &Molecular Medicine, College of Medicine, Chosun University, Gwangju 61452, Republic of Korea.
authors.forename: Ra Young
authors.initials: RY
authors.lastname: Park
authors.affiliation: Department of Anatomy, School of Medicine, Jeju National University, Jeju-Do 690-756, Republic of Korea.
authors.forename: Sang Pil
authors.initials: SP
authors.lastname: Yoon
authors.affiliation: Diatech Korea Co, Ltd, Saemal-ro 5-gil, Songpa-gu, Seoul 05807, Republic of Korea.
authors.forename: Gum-Yong
authors.initials: GY
authors.lastname: Kang
authors.affiliation: Diatech Korea Co, Ltd, Saemal-ro 5-gil, Songpa-gu, Seoul 05807, Republic of Korea.
authors.forename: Je-Hyun
authors.initials: JH
authors.lastname: Baek
authors.affiliation: Division of Rheumatology, Immunology and Allergy, Brigham and Women's Hospital, Smith652, One Jimmy Fund Way, Boston, Massachusetts 02115, USA.
authors.forename: Paul
authors.initials: P
authors.lastname: Anderson
authors.affiliation: Department of Cell Biology, Microbiology, and Molecular Biology, College of Arts and Sciences, University of South Florida, Tampa, Florida 33620, USA.
authors.forename: Younghoon
authors.initials: Y
authors.lastname: Kee
authors.affiliation: Department of Cellular &Molecular Medicine, College of Medicine, Chosun University, Gwangju 61452, Republic of Korea.
authors.forename: Takbum
authors.initials: T
authors.lastname: Ohn
mesh_terms: D018053:Arsenites / Q000633:toxicity*; D045744:Cell Line, Tumor; D003594:Cytoplasmic Granules / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D008239:Lysine / Q000378:metabolism; D000075687:NEDD8 Protein / Q000235:genetics* / Q000378:metabolism; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D018384:Oxidative Stress; D011132:Polyribosomes / Q000187:drug effects; D014176:Protein Biosynthesis / Q000187:drug effects*; D012261:Ribonucleoproteins / Q000235:genetics / Q000378:metabolism; D000068103:Serine-Arginine Splicing Factors / Q000235:genetics* / Q000378:metabolism; D044763:Ubiquitin-Conjugating Enzymes / Q000235:genetics / Q000378:metabolism
chemical_list: D018053:Arsenites; D000075687:NEDD8 Protein; C082860:NEDD8 protein, human; D012261:Ribonucleoproteins; C491388:SRSF3 protein, human; C026452:messenger ribonucleoprotein; D000068103:Serine-Arginine Splicing Factors; D044763:Ubiquitin-Conjugating Enzymes; C000596156:UBE2M protein, human; D008239:Lysine; C015001:arsenite