Interaction of the Vp3 nuclear localization signal with the importin alpha 2/beta heterodimer directs nuclear entry of infecting simian virus 40.
For nuclear entry of large nucleoprotein complexes, it is thought that one key nuclear localization signal (NLS) of a protein component becomes exposed to mediate importin recognition. We show that the nuclear entry of simian virus 40 involves a dynamic interplay between two distinct interiorly situated capsid NLSs, the Vp1 NLS and the Vp3 NLS, and the selective exposure and importin recognition of the Vp3 NLS. The Vp3 NLS-null mutants assembled normally into virion-like particles (VLP) in mutant DNA-transfected cells. When used to infect a new host, the null VLP entered the cell normally but was impaired in viral DNA nuclear entry due to a lack of recognition by the importin alpha 2/beta heterodimer, leading to reduced viability. Both Vp3 and Vp1 NLSs directed importin interaction in vitro, but the Vp1 NLS, which overlaps the Vp1 DNA binding domain, did not bind importins in the presence of DNA. The results suggest that certain canonical NLSs within a nucleoprotein complex, such as the Vp1 NLS, can be masked from functioning by binding to the nucleic acid component and that the availability of an NLS that is not masked and can become exposed for importin binding, such as the Vp3 NLS, is a general feature of the nuclear entry of the nucleoprotein complexes, including those of other animal viruses.
Department of Molecular, Cell, and Developmental Biology and Molecular Biology Institute, University of California at Los Angeles, Los Angeles, California 90095, USA.
Akira
A
Nakanishi
Dorothy
D
Shum
Hiroshi
H
Morioka
Eiko
E
Otsuka
Harumi
H
Kasamatsu
D000595:Amino Acid Sequence; D000818:Animals; D002213:Capsid / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D036022:Capsid Proteins; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism / Q000821:virology; D004261:DNA Replication; D004279:DNA, Viral / Q000378:metabolism; D019281:Dimerization; D008969:Molecular Sequence Data; D009154:Mutation; D019913:Nuclear Localization Signals / Q000235:genetics / Q000378:metabolism*; D013539:Simian virus 40 / Q000502:physiology*; D014771:Virion / Q000378:metabolism; D014779:Virus Replication; D028901:alpha Karyopherins / Q000378:metabolism*; D028961:beta Karyopherins / Q000378:metabolism*
D036022:Capsid Proteins; D004279:DNA, Viral; D019913:Nuclear Localization Signals; C062481:VP1 protein, polyomavirus; C085902:VP3 protein, Simian virus 40; D028901:alpha Karyopherins; D028961:beta Karyopherins