Synthesis and assembly of virus-like particles of human papillomaviruses type 6 and type 16 in fission yeast Schizosaccharomyces pombe.
We have synthesized capsid proteins of human papillomavirus types 6 (HPV 6) and 16 (HPV 16) in fission yeast Schizosaccharomyces pombe and produced virus-like particles (VLP). The capsid proteins were localized in the nucleus by indirect immunofluorescence and cell fractionation analyses. The VLP were produced in both yeast clones synthesizing L1 alone and L/L2 and purified by sulfato-cellulofine chromatography. Electron microscopic examination showed that these VLP were similar in structure to native HPV particles. Two HPV 16 L1 variants (16 B27L1 and 16 T3L1), isolated from benign cervical samples, produced many more (68- and 14-fold) VLP than the prototype L1 (16 PL1) derived from cervical carcinoma. Coexpression of the HPV 6 L2 protein with 6 L1 and 16 B27L1 proteins increased the production level of the VLP four- and twofold, respectively. The L2 was not detected in the VLP purified with sulfato-cellulofine column, although the L2 was purified in the same fraction containing HPV 6 and 16 B27-VLP by size-fractionation using Sepharose column. Interaction between 6 L2 and 6/16 L1 proteins was not detected by the coimmunoprecipitation assays with either L1 or L2 antibodies. These results suggest that the L2 is not incorporated into the VLP synthesized in yeast.
Sasagawa
T
T
Imperial Cancer Research Fund Tumour Virus Group, Department of Pathology, Cambridge, United Kingdom.
Pushko
P
P
Steers
G
G
Gschmeissner
S E
SE
Hajibagheri
M A
MA
Finch
J
J
Crawford
L
L
Tommasino
M
M
D001483:Base Sequence; D002213:Capsid / Q000235:genetics*; D036022:Capsid Proteins; D003001:Cloning, Molecular; D017931:DNA Primers; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D009856:Oncogene Proteins, Viral / Q000235:genetics*; D027383:Papillomaviridae / Q000502:physiology* / Q000648:ultrastructure; D011994:Recombinant Proteins / Q000235:genetics; D012568:Schizosaccharomyces / Q000235:genetics*; D014764:Viral Proteins; D014771:Virion / Q000302:isolation & purification / Q000502:physiology*
D036022:Capsid Proteins; D017931:DNA Primers; D009856:Oncogene Proteins, Viral; D011994:Recombinant Proteins; D014764:Viral Proteins; C091560:L1 protein, Human papillomavirus type 6; C063317:L1 protein, Human papillomavirus type 16