Imaging and characterizing influenza A virus mRNA transport in living cells.
The mechanisms of influenza A virus mRNA intracellular transport are still not clearly understood. Here, we visualized the distribution and transport of influenza A virus mRNA in living cells using molecular beacon (MB) technology. Confocal-FRAP measurements determined that the transport of influenza A virus intronless mRNA, in both nucleus and cytoplasm, was energy dependent, being similar to that of Poly(A)(+) RNA. Drug inhibition studies in living cells revealed that the export of influenza A virus mRNA is independent of the CRM1 pathway, while the function of RNA polymerase II (RNAP-II) may be needed. In addition, viral NS1 protein and cellular TAP protein were found associated with influenza A virus mRNA in the cell nucleus. These findings characterize influenza A virus mRNA transport in living cells and suggest that influenza A virus mRNA may be exported from the nucleus by the cellular TAP/p15 pathway with NS1 protein and RNAP-II participation.
Wang
Wei
W
State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, China.
Cui
Zong-Qiang
ZQ
Han
Han
H
Zhang
Zhi-Ping
ZP
Wei
Hong-Ping
HP
Zhou
Ya-Feng
YF
Chen
Ze
Z
Zhang
Xian-En
XE
D000818:Animals; D002460:Cell Line; D002467:Cell Nucleus / Q000737:chemistry / Q000821:virology; D003593:Cytoplasm / Q000821:virology; D003609:Dactinomycin / Q000494:pharmacology; D004285:Dogs; D036681:Fluorescence Recovery After Photobleaching; D047468:Immunoprecipitation; D009980:Influenza A virus / Q000235:genetics / Q000378:metabolism*; D028884:Karyopherins / Q000378:metabolism; D018613:Microscopy, Confocal; D029543:Nucleocytoplasmic Transport Proteins / Q000032:analysis; D034443:RNA Transport / Q000187:drug effects; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism*; D012367:RNA, Viral / Q000032:analysis / Q000378:metabolism*; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism; D017361:Viral Nonstructural Proteins / Q000032:analysis; D000097606:Exportin 1 Protein
D003609:Dactinomycin; D028884:Karyopherins; D029543:Nucleocytoplasmic Transport Proteins; D012333:RNA, Messenger; D012367:RNA, Viral; D018160:Receptors, Cytoplasmic and Nuclear; D017361:Viral Nonstructural Proteins; D000097606:Exportin 1 Protein; C085820:INS1 protein, influenza virus