DNA packaging by L1 and L2 capsid proteins of bovine papillomavirus type 1.
Encapsidation of circular DNA by papillomavirus capsid protein was investigated in Cos-1 cells. Plasmids carrying both an SV40 origin of replication (ori) and an E. coli ori were introduced into Cos-1 cells by DNA transfection PV capsid proteins were supplied in trans by recombinant vaccinia viruses. Pseudovirions were purified from infected cells and their packaged DNA was extracted and used to transform E. coli as an indication of packaging efficacy. VLPs assembled from BPV-1 L1 alone packaged little plasmid DNA, whereas VLPs assembled from BPV-1 L1 + L2 packaged plasmid DNA at least 50 times more effectively. BPV-1 L1 + L2 VLPs packaged a plasmid containing BPV-1 sequence 8.2 +/- 3.1 times more effectively than a plasmid without BPV sequences. Using a series of plasmid constructs comprising a core BPV-1 sequence and spacer DNA it was demonstrated that BPV VLPs could accommodate a maximum of about 10.2 kb of plasmid DNA, and that longer closed circular DNA was truncated to produce less dense virions with shorter plasmid sequences. The present study suggests that packaging of genome within PV virions involves interaction of L2 protein with specific DNA sequences, and demonstrates that PV pseudovirions have the potential to be used as DNA delivery vectors for plasmids of up to 10.2 kb.
Zhao
K N
KN
Centre for Immunology and Cancer Research, University of Queensland, Princess Alexandra Hospital, Woolloongabba, Australia.
Sun
X Y
XY
Frazer
I H
IH
Zhou
J
J
D000818:Animals; D052688:Bovine papillomavirus 1 / Q000378:metabolism* / Q000502:physiology; D002213:Capsid / Q000378:metabolism*; D036022:Capsid Proteins; D002417:Cattle; D004247:DNA / Q000378:metabolism*; D009690:Nucleic Acid Conformation; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013329:Structure-Activity Relationship; D014771:Virion; D019065:Virus Assembly
D036022:Capsid Proteins; C085217:L1 protein, Bovine papillomavirus; C085218:L2 protein, Bovine papillomavirus; D011993:Recombinant Fusion Proteins; D004247:DNA