Nucleoporin 153 arrests the nuclear import of hepatitis B virus capsids in the nuclear basket.
Virtually all DNA viruses including hepatitis B viruses (HBV) replicate their genome inside the nucleus. In non-dividing cells, the genome has to pass through the nuclear pore complexes (NPCs) by the aid of nuclear transport receptors as e.g. importin beta (karyopherin). Most viruses release their genome in the cytoplasm or at the cytosolic face of the NPC, as the diameter of their capsids exceeds the size of the NPC. The DNA genome of HBV is derived from reverse transcription of an RNA pregenome. Genome maturation occurs in cytosolic capsids and progeny capsids can deliver the genome into the nucleus causing nuclear genome amplification. The karyophilic capsids are small enough to pass the NPC, but nuclear entry of capsids with an immature genome is halted in the nuclear basket on the nuclear side of the NPC, and the genome remains encapsidated. In contrast, capsids with a mature genome enter the basket and consequently liberate the genome. Investigating the difference between immature and mature capsids, we found that mature capsids had to disintegrate in order to leave the nuclear basket. The arrest of a karyophilic cargo at the nuclear pore is a rare phenomenon, which has been described for only very few cellular proteins participating in nuclear entry. We analyzed the interactions causing HBV capsid retention. By pull-down assays and partial siRNA depletion, we showed that HBV capsids directly interact with nucleoporin 153 (Nup153), an essential protein of the nuclear basket which participates in nuclear transport via importin beta. The binding sites of importin beta and capsids were shown to overlap but capsid binding was 150-fold stronger. In cellulo experiments using digitonin-permeabilized cells confirmed the interference between capsid binding and nuclear import by importin beta. Collectively, our findings describe a unique nuclear import strategy not only for viruses but for all karyophilic cargos.
Schmitz
André
A
Institute of Medical Virology, Justus Liebig University, Giessen, Germany.
Schwarz
Alexandra
A
Foss
Michael
M
Zhou
Lixin
L
Rabe
Birgit
B
Hoellenriegel
Julia
J
Stoeber
Miriam
M
Panté
Nelly
N
Kann
Michael
M
D021581:Active Transport, Cell Nucleus / Q000502:physiology*; D000818:Animals; D036022:Capsid Proteins / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism* / Q000821:virology; D006367:HeLa Cells; D006515:Hepatitis B virus / Q000378:metabolism*; D006801:Humans; D047468:Immunoprecipitation; D028861:Nuclear Pore Complex Proteins / Q000378:metabolism*; D034741:RNA, Small Interfering; D014779:Virus Replication / Q000502:physiology*; D014982:Xenopus laevis
D036022:Capsid Proteins; D028861:Nuclear Pore Complex Proteins; D034741:RNA, Small Interfering