[en] Available data suggest that the Varicella-Zoster virus (VZV) IE4 protein acts as an important regulator on VZV and cellular genes expression and could exert its functions at post-transcriptional level. However, the molecular mechanisms supported by this protein are not yet fully characterized. In the present study, we have attempted to clarify this IE4-mediated gene regulation and identify some cellular partners of IE4. By yeast two-hybrid and immunoprecipitation analysis, we showed that IE4 interacts with three shuttling SR proteins, namely ASF/SF2, 9G8 and SRp20. We positioned the binding domain in the IE4 RbRc region and we showed that these interactions are not bridged by RNA. We demonstrated also that IE4 strongly interacts with the main SR protein kinase, SRPK1, and is phosphorylated in in vitro kinase assay on residue Ser-136 contained in the Rb domain. By Northwestern analysis, we showed that IE4 is able to bind RNA through its arginine-rich region and in immunoprecipitation experiments the presence of RNA stabilizes complexes containing IE4 and the cellular export factors TAP/NXF1 and Aly/REF since the interactions are RNase-sensitive. Finally, we determined that IE4 influences the export of reporter mRNAs and clearly showed, by TAP/NXF1 knockdown, that VZV infection requires the TAP/NXF1 export pathway to express some viral transcripts. We thus highlighted a new example of viral mRNA export factor and proposed a model of IE4-mediated viral mRNAs export.
Research Center/Unit :
GIGA-I3 - Giga-Infection, Immunity and Inflammation - ULiège
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Ote, Isabelle ; Université de Liège - ULiège > Département des Sciences de la Vie > GIGA-R: Virologie et Immunologie
Vandevenne, Patricia ; Université de Liège - ULiège > Département des sciences de la vie > GIGA-R : Virologie et immunologie
Bontems, Sébastien ; Université de Liège - ULiège > Département des Sciences de la Vie > GIGA-R: Virologie et Immunologie
Medina-Palazon, C.; INSERM U758, ENS-Lyon, Lyon, France > Laboratoire de Virologie Humaine,
Manet, E.; INSERM U758, ENS-Lyon, Lyon, France > Laboratoire de Virologie Humaine,
Piette, Jacques ; Université de Liège - ULiège > Département des sciences de la vie > GIGA-R : Virologie - Immunologie - Département des sciences de la vie - GIGA-Research
Lebrun, Marielle ; Université de Liège - ULiège > Département des sciences de la vie > GIGA-R : Virologie et immunologie
Sadzot-Delvaux, Catherine ; Université de Liège - ULiège > Département des sciences de la vie > GIGA-R : Virologie et immunologie
Language :
English
Title :
Varicella-Zoster Virus IE4 Protein Interacts with SR Proteins and Exports mRNAs through the TAP/NXF1 Pathway.
Publication date :
18 November 2009
Journal title :
PLoS ONE
eISSN :
1932-6203
Publisher :
Public Library of Science, San Franscisco, United States - California
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