Sequence determination of human papillomavirus type 6a and assembly of virus-like particles in Saccharomyces cerevisiae.
Human papillomavirus 6a (HPV6a), the most abundant HPV6 subtype, was detected in a vulvar condyloma acuminatum. The complete genome of HPV6a was cloned, and its DNA sequence was shown to be over 97% identical to the HPV6b sequence. Of the eight open reading frames (ORFs) of HPV6a, only the imputed amino acid sequence of the major capsid protein L1 was identical to the corresponding HPV6b sequence; all other HPV6a ORFs showed amino acid changes compared to the HPV6b ORFs. The HPV6a L1 or the L1 + L2 ORFs were expressed in the yeast Saccharomyces cerevisiae. Self-assembly of the L1 capsid protein into virus-like particles (VLPs) was demonstrated both in the L1 as well as L1 + L2 coexpressing yeast strains. Copurification of the L1 and L2 proteins showed complex formation of the L1 and L2 proteins in the yeast-derived VLPs of coexpressing strains.
Hofmann
K J
KJ
Department of Virus and Cell Biology, Merck Research Laboratories, West Point, Pennsylvania 19486, USA.
Cook
J C
JC
Joyce
J G
JG
Brown
D R
DR
Schultz
L D
LD
George
H A
HA
Rosolowsky
M
M
Fife
K H
KH
Jansen
K U
KU
D000328:Adult; D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D003062:Codon / Q000235:genetics; D003218:Condylomata Acuminata / Q000821:virology; D017931:DNA Primers; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification; D005260:Female; D014644:Genetic Variation; D016679:Genome, Viral; D006801:Humans; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D027383:Papillomaviridae / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D011644:Puerperal Disorders / Q000821:virology; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae; D014845:Vulvar Diseases / Q000821:virology
D003062:Codon; D017931:DNA Primers; D004279:DNA, Viral