Immunosuppressive Yersinia Effector YopM Binds DEAD Box Helicase DDX3 to Control Ribosomal S6 Kinase in the Nucleus of Host Cells.
Yersinia outer protein M (YopM) is a crucial immunosuppressive effector of the plaque agent Yersinia pestis and other pathogenic Yersinia species. YopM enters the nucleus of host cells but neither the mechanisms governing its nucleocytoplasmic shuttling nor its intranuclear activities are known. Here we identify the DEAD-box helicase 3 (DDX3) as a novel interaction partner of Y. enterocolitica YopM and present the three-dimensional structure of a YopM:DDX3 complex. Knockdown of DDX3 or inhibition of the exportin chromosomal maintenance 1 (CRM1) increased the nuclear level of YopM suggesting that YopM exploits DDX3 to exit the nucleus via the CRM1 export pathway. Increased nuclear YopM levels caused enhanced phosphorylation of Ribosomal S6 Kinase 1 (RSK1) in the nucleus. In Y. enterocolitica infected primary human macrophages YopM increased the level of Interleukin-10 (IL-10) mRNA and this effect required interaction of YopM with RSK and was enhanced by blocking YopM's nuclear export. We propose that the DDX3/CRM1 mediated nucleocytoplasmic shuttling of YopM determines the extent of phosphorylation of RSK in the nucleus to control transcription of immunosuppressive cytokines.
Institute of Medical Microbiology, Virology and Hygiene, University Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany.
Laura
L
Berneking
Institute of Medical Microbiology, Virology and Hygiene, University Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany.
Marie
M
Schnapp
Institute of Medical Microbiology, Virology and Hygiene, University Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany.
Andreas
A
Rumm
Institute of Medical Microbiology, Virology and Hygiene, University Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany.
Claudia
C
Trasak
Institute of Medical Microbiology, Virology and Hygiene, University Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany.
Klaus
K
Ruckdeschel
Bioinformatics Core, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.|Heinrich-Pette-Institute (HPI), Leibniz Institute for Experimental Virology, Research Group Virus Genomics, Hamburg, Germany.
Malik
M
Alawi
Heinrich-Pette-Institute (HPI), Leibniz Institute for Experimental Virology, Research Group Virus Genomics, Hamburg, Germany.
Adam
A
Grundhoff
European Molecular Biology Laboratory (EMBL), Hamburg Outstation, Hamburg, Germany.
Alexey G
AG
Kikhney
Institute of Immunology, University Medical Center, Hamburg, Germany.
Friedrich
F
Koch-Nolte
Institute of Clinical Chemistry, University Medical Center, Hamburg, Germany.
Friedrich
F
Buck
Institute of Medical Microbiology, Virology and Hygiene, University Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany.|Institute of Biochemistry and Molecular Biology, University of Hamburg, Laboratory of Structural Biology of Infection and Inflammation, Hamburg, Germany.|The Hamburg Centre for Ultrafast Imaging, Hamburg, Germany.
Markus
M
Perbandt
Institute of Biochemistry and Molecular Biology, University of Hamburg, Laboratory of Structural Biology of Infection and Inflammation, Hamburg, Germany.
Christian
C
Betzel
European Molecular Biology Laboratory (EMBL), Hamburg Outstation, Hamburg, Germany.
Dmitri I
DI
Svergun
Institute of Medical Microbiology, Virology and Hygiene, University Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany.
Moritz
M
Hentschke
Institute of Medical Microbiology, Virology and Hygiene, University Medical Center Hamburg-Eppendorf (UKE), Hamburg, Germany.
Martin
M
Aepfelbacher
D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D015153:Blotting, Western; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D018360:Crystallography, X-Ray; D053487:DEAD-box RNA Helicases / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D005786:Gene Expression Regulation / Q000502:physiology*; D059014:High-Throughput Nucleotide Sequencing; D006790:Host-Parasite Interactions / Q000502:physiology; D006801:Humans; D007108:Immune Tolerance / Q000502:physiology; D047468:Immunoprecipitation; D008264:Macrophages / Q000382:microbiology; D013058:Mass Spectrometry; D018613:Microscopy, Confocal; D016133:Polymerase Chain Reaction; D021381:Protein Transport / Q000502:physiology; D038744:Ribosomal Protein S6 Kinases, 90-kDa / Q000096:biosynthesis*; D037521:Virulence Factors / Q000276:immunology / Q000378:metabolism; D015009:Yersinia Infections / Q000276:immunology* / Q000378:metabolism; D015008:Yersinia enterocolitica
D001425:Bacterial Outer Membrane Proteins; D037521:Virulence Factors; C060772:yopM protein, Yersinia; C069329:RPS6KA1 protein, human; D038744:Ribosomal Protein S6 Kinases, 90-kDa; C119332:DDX3X protein, human; D053487:DEAD-box RNA Helicases