title: Inhibition of eukaryotic translation by tetratricopeptide-repeat proteins of Orientia tsutsugamushi.
abstract: Orientia tsutsugamushi, an obligate intracellular bacterium, is the causative agent of scrub typhus. The genome of Orientia tsutsugamushi has revealed multiple ORFs encoding tetratricopeptide-repeat (TPR) proteins. The TPR protein family has been shown to be involved in a diverse spectrum of cellular functions such as cell cycle control, transcription, protein transport, and protein folding, especially in eukaryotic cells. However, little is known about the function of the TPR proteins in O. tsutsugamushi. To investigate the potential role of TPR proteins in host-pathogen interaction, two oriential TPR proteins were expressed in E. coli and applied for GSTpull down assay. DDX3, a DEAD-box containing RNA helicase, was identified as a specific eukaryotic target of the TPR proteins. Since the RNA helicase is involved in multiple RNA-modifying processes such as initiation of translation reaction, we performed in vitro translation assay in the presence of GST-TPR fusion proteins by using rabbit reticulocyte lysate system. The TPR proteins inhibited in vitro translation of a reporter luciferase in a dose dependent manner whereas the GST control proteins did not. These results suggested TPR proteins of O. tsutsugamushi might be involved in the modulation of eukaryotic translation through the interaction with DDX3 RNA helicase after secretion into host cytoplasm.
authors.affiliation: Department of Microbiology and Immunology, Seoul National University College of Medicine, Seoul, 03080, Republic of Korea.
authors.forename: Sunyoung
authors.initials: S
authors.lastname: Bang
authors.affiliation: Department of Microbiology and Immunology, Seoul National University College of Medicine, Seoul, 03080, Republic of Korea.|Department of Biomedical Science, Seoul National University College of Medicine, Seoul, 03080, Republic of Korea.
authors.forename: Chan-Ki
authors.initials: CK
authors.lastname: Min
authors.affiliation: Department of Microbiology and Immunology, Seoul National University College of Medicine, Seoul, 03080, Republic of Korea.|Department of Biomedical Science, Seoul National University College of Medicine, Seoul, 03080, Republic of Korea.
authors.forename: Na-Young
authors.initials: NY
authors.lastname: Ha
authors.affiliation: Department of Microbiology and Immunology, Seoul National University College of Medicine, Seoul, 03080, Republic of Korea.
authors.forename: Myung-Sik
authors.initials: MS
authors.lastname: Choi
authors.affiliation: Department of Microbiology and Immunology, Seoul National University College of Medicine, Seoul, 03080, Republic of Korea.
authors.forename: Ik-Sang
authors.initials: IS
authors.lastname: Kim
authors.affiliation: Divisions of Infectious Diseases, Chungnam National University Hospital, Daejeon, 35015, Republic of Korea. idalicekim@gmail.com.
authors.forename: Yeon-Sook
authors.initials: YS
authors.lastname: Kim
authors.affiliation: Department of Microbiology and Immunology, Seoul National University College of Medicine, Seoul, 03080, Republic of Korea. chonh@snu.ac.kr.|Department of Biomedical Science, Seoul National University College of Medicine, Seoul, 03080, Republic of Korea. chonh@snu.ac.kr.|Institute of Endemic Disease, Seoul National University Medical Research Center and Bundang Hospital, Seoul, 35015, Republic of Korea. chonh@snu.ac.kr.
authors.forename: Nam-Hyuk
authors.initials: NH
authors.lastname: Cho
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000302:isolation & purification / Q000378:metabolism*; D002460:Cell Line; D002474:Cell-Free System; D053487:DEAD-box RNA Helicases / Q000378:metabolism; D054884:Host-Pathogen Interactions; D006801:Humans; D012285:Orientia tsutsugamushi / Q000737:chemistry*; D011485:Protein Binding; D014176:Protein Biosynthesis / Q000187:drug effects*; D011500:Protein Synthesis Inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D011817:Rabbits
chemical_list: D001426:Bacterial Proteins; D011500:Protein Synthesis Inhibitors; C119332:DDX3X protein, human; D053487:DEAD-box RNA Helicases
keywords: DDX3; Orientia tsutsugamushi; tetratricopeptide proteins; translation
references.citation: Science. 1997 Mar 7;275(5305):1468-71
references.pmid: 9045610
references.citation: Infect Immun. 2006 Jul;74(7):4246-53
references.pmid: 16790799
references.citation: J Struct Biol. 2001 May-Jun;134(2-3):117-31
references.pmid: 11551174
references.citation: Nucleic Acids Res. 2006;34(15):4168-80
references.pmid: 16936318
references.citation: PLoS One. 2014 Aug 28;9(8):e105652
references.pmid: 25166298
references.citation: J Bacteriol. 2015 Oct;197(19):3097-109
references.pmid: 26170417
references.citation: DNA Res. 2008 Aug;15(4):185-99
references.pmid: 18508905
references.citation: Proteomics. 2010 Apr;10(8):1699-715
references.pmid: 20186754
references.citation: Proc Natl Acad Sci U S A. 2007 May 8;104(19):7981-6
references.pmid: 17483455
references.citation: PLoS Biol. 2005 Aug;3(8):e248
references.pmid: 15984913
references.citation: J Mol Biol. 2007 Sep 7;372(1):150-9
references.pmid: 17631897
references.citation: PLoS Negl Trop Dis. 2015 Mar 13;9(3):e0003585
references.pmid: 25768004
references.citation: PLoS Negl Trop Dis. 2015 Aug 14;9(8):e0003971
references.pmid: 26274584
references.citation: Am J Trop Med Hyg. 1982 May;31(3 Pt 1):532-40
references.pmid: 6805348
references.citation: Infect Immun. 2000 Feb;68(2):594-602
references.pmid: 10639422
references.citation: Nat Rev Mol Cell Biol. 2004 Mar;5(3):232-41
references.pmid: 14991003
references.citation: J Virol. 2007 Dec;81(24):13922-6
references.pmid: 17855521
references.citation: Comp Funct Genomics. 2008;:623145
references.pmid: 18528528
references.citation: Trends Genet. 2007 Oct;23(10):511-20
references.pmid: 17822801
references.citation: Bioessays. 1999 Nov;21(11):932-9
references.pmid: 10517866
references.citation: Mol Microbiol. 2004 Feb;51(4):987-1001
references.pmid: 14763975
references.citation: PLoS Genet. 2006 May;2(5):e76
references.pmid: 16703114
references.citation: Infect Immun. 2001 Mar;69(3):1265-72
references.pmid: 11179287
references.citation: Cell. 2004 Oct 29;119(3):381-92
references.pmid: 15507209
references.citation: Trends Parasitol. 2006 Feb;22(2):60-5
references.pmid: 16406333
references.citation: Curr Opin Infect Dis. 2003 Oct;16(5):429-36
references.pmid: 14501995
references.citation: Gene. 2006 Feb 15;367:17-37
references.pmid: 16337753
references.citation: J Biol Chem. 1999 May 28;274(22):15751-6
references.pmid: 10336476