title: Nuclear import of HPV11 L1 capsid protein is mediated by karyopherin alpha2beta1 heterodimers.
abstract: L1 major capsid proteins of human papillomaviruses (HPVs) enter the nuclei of host cells at two times during the viral life cycle: 1) after infection and 2) later during the productive phase, when they assemble the replicated HPV genomic DNA into infectious virions. L1 proteins are stable in two oligomeric configurations: as homopentameric capsomers, and as capsids composed of 72 capsomers. We found that intact L1 capsids of HPV type 11 cannot enter the nucleus, suggesting that capsid disassembly may be required for HPV11 L1 nuclear import. We established that HPV11 L1 is imported in a receptor-mediated manner into the nuclei of digitonin-permeabilized HeLa cells. HPV11 L1 docked at the nuclear pore complexes via karyopherin alpha2beta1 heterodimers. Anti-karyopherin-beta1 and anti-karyopherin alpha2 antibodies specifically inhibited nuclear import of HPV11 L1. Moreover, nuclear import of HPV11 L1 could be reconstituted using karyopherin alpha2, beta1, RanGDP and p10. In agreement with the docking and import data, we found that HPV11 L1 binds to karyopherin alpha2 and that this interaction is inhibited by a peptide representing the classical nuclear localization signal of SV40 T antigen. These results strongly suggest that HPV11 L1 enters the nucleus of the infected host cell via the karyopherin alpha2beta1 pathway.
authors.affiliation: Laboratory of Cell Biology, Biology Department, Boston College, Chestnut Hill, Massachusetts 02467, USA.
authors.forename: E
authors.initials: E
authors.lastname: Merle
authors.forename: R C
authors.initials: RC
authors.lastname: Rose
authors.forename: L
authors.initials: L
authors.lastname: LeRoux
authors.forename: J
authors.initials: J
authors.lastname: Moroianu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001693:Biological Transport, Active; D002213:Capsid / Q000378:metabolism*; D036022:Capsid Proteins; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism / Q000821:virology; D019281:Dimerization; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D019913:Nuclear Localization Signals; D009687:Nuclear Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D009856:Oncogene Proteins, Viral / Q000378:metabolism*; D027383:Papillomaviridae / Q000378:metabolism; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D028901:alpha Karyopherins; D028961:beta Karyopherins
chemical_list: D036022:Capsid Proteins; D002352:Carrier Proteins; C094776:L1 protein, Human papillomavirus type 11; D019913:Nuclear Localization Signals; D009687:Nuclear Proteins; D009856:Oncogene Proteins, Viral; D011994:Recombinant Proteins; D028901:alpha Karyopherins; D028961:beta Karyopherins; C087978:karyopherin alpha 2