Dephosphorylation of HuR protein during alphavirus infection is associated with HuR relocalization to the cytoplasm.
We have demonstrated previously that the cellular HuR protein binds U-rich elements in the 3' untranslated region (UTR) of Sindbis virus RNA and relocalizes from the nucleus to the cytoplasm upon Sindbis virus infection in 293T cells. In this study, we show that two alphaviruses, Ross River virus and Chikungunya virus, lack the conserved high-affinity U-rich HuR binding element in their 3' UTRs but still maintain the ability to interact with HuR with nanomolar affinities through alternative binding elements. The relocalization of HuR protein occurs during Sindbis infection of multiple mammalian cell types as well as during infections with three other alphaviruses. Interestingly, the relocalization of HuR is not a general cellular reaction to viral infection, as HuR protein remained largely nuclear during infections with dengue and measles virus. Relocalization of HuR in a Sindbis infection required viral gene expression, was independent of the presence of a high-affinity U-rich HuR binding site in the 3' UTR of the virus, and was associated with an alteration in the phosphorylation state of HuR. Sindbis virus-induced HuR relocalization was mechanistically distinct from the movement of HuR observed during a cellular stress response, as there was no accumulation of caspase-mediated HuR cleavage products. Collectively, these data indicate that virus-induced HuR relocalization to the cytoplasm is specific to alphavirus infections and is associated with distinct posttranslational modifications of this RNA-binding protein.
Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, Colorado 80523, USA.
Alexa M
AM
Dickson
John R
JR
Anderson
Michael D
MD
Barnhart
Kevin J
KJ
Sokoloski
Lauren
L
Oko
Mateusz
M
Opyrchal
Evanthia
E
Galanis
Carol J
CJ
Wilusz
Thomas E
TE
Morrison
Jeffrey
J
Wilusz
D020413:3' Untranslated Regions / Q000502:physiology; D000524:Alphavirus / Q000235:genetics / Q000378:metabolism*; D018354:Alphavirus Infections / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D020169:Caspases / Q000235:genetics / Q000378:metabolism; D002522:Chlorocebus aethiops; D003593:Cytoplasm / Q000235:genetics / Q000378:metabolism* / Q000821:virology; D051959:ELAV Proteins / Q000235:genetics / Q000378:metabolism*; D015967:Gene Expression Regulation, Viral / Q000502:physiology; D057809:HEK293 Cells; D006801:Humans; D010766:Phosphorylation / Q000235:genetics; D011499:Protein Processing, Post-Translational; D021381:Protein Transport / Q000235:genetics; D059748:Proteolysis; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism; D014709:Vero Cells
D020413:3' Untranslated Regions; D051959:ELAV Proteins; D012367:RNA, Viral; D020169:Caspases