Possible role for cellular karyopherins in regulating polyomavirus and papillomavirus capsid assembly.
Polyomavirus and papillomavirus (papovavirus) capsids are composed of 72 capsomeres of their major capsid proteins, VP1 and L1, respectively. After translation in the cytoplasm, L1 and VP1 pentamerize into capsomeres and are then imported into the nucleus using the cellular alpha and beta karyopherins. Virion assembly only occurs in the nucleus, and cellular mechanisms exist to prevent premature capsid assembly in the cytosol. We have identified the karyopherin family of nuclear import factors as possible "chaperones" in preventing the cytoplasmic assembly of papovavirus capsomeres. Recombinant murine polyomavirus (mPy) VP1 and human papillomavirus type 11 (HPV11) L1 capsomeres bound the karyopherin heterodimer alpha2beta1 in vitro in a nuclear localization signal (NLS)-dependent manner. Because the amino acid sequence comprising the NLS of VP1 and L1 overlaps the previously identified DNA binding domain, we examined the relationship between karyopherin and DNA binding of both mPy VP1 and HPV11 L1. Capsomeres of L1, but not VP1, bound by karyopherin alpha2beta1 or beta1 alone were unable to bind DNA. VP1 and L1 capsomeres could bind both karyopherin alpha2 and DNA simultaneously. Both VP1 and L1 capsomeres bound by karyopherin alpha2beta1 were unable to assemble into capsids, as shown by in vitro assembly reactions. These results support a role for karyopherins as chaperones in the in vivo regulation of viral capsid assembly.
Bird
Gregory
G
Department of Pediatrics, University of Colorado School of Medicine, UCHSC at Fitzsimons RC1-North, Room 4100, Mail Stop 8302, P.O. Box 6511, Aurora, CO 80045, USA.
O'Donnell
Malinda
M
Moroianu
Junona
J
Garcea
Robert L
RL
D000818:Animals; D002213:Capsid / Q000378:metabolism*; D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D052140:Human papillomavirus 11 / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure*; D006801:Humans; D051379:Mice; D018832:Molecular Chaperones / Q000235:genetics / Q000378:metabolism; D009856:Oncogene Proteins, Viral / Q000235:genetics / Q000378:metabolism; D011120:Polyomavirus / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure*; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D019065:Virus Assembly; D028901:alpha Karyopherins / Q000235:genetics / Q000378:metabolism*; D028961:beta Karyopherins / Q000235:genetics / Q000378:metabolism*
D036022:Capsid Proteins; C094776:L1 protein, Human papillomavirus type 11; D018832:Molecular Chaperones; D009856:Oncogene Proteins, Viral; D011993:Recombinant Fusion Proteins; C062481:VP1 protein, polyomavirus; D028901:alpha Karyopherins; D028961:beta Karyopherins; D004247:DNA