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PMID: 8386271 Published · ppublish English Journal Article

Characterization of human papillomavirus type 11 E1 and E2 proteins expressed in insect cells.

Journal of virology ·Vol. 67 ·No. 5 ·1993-05-00 ·Pages 2655-63

Bream GL, Ohmstede CA, Phelps WC

Abstract

The study of human papillomavirus replication has been hampered by the lack of an in vitro system which reliably supports virus replication. Recent results from the bovine papillomavirus (BPV) system indicate that the E1 and E2 proteins are the only viral gene products required for replication. By analogy with simian virus 40 large T antigen, E1 is thought to possess ATPase and helicase activity, which may play a direct role in viral DNA replication. The precise role of E2 is unclear, but it may function in part to help localize E1 to the replication origin. We have initiated a study of replication in the human papillomavirus type 11 system which, by analogy to BPV, has focused on the E1 and E2 proteins of this virus. We have expressed the full-length E1 and E2 proteins in Sf9 insect cells by using a baculovirus expression vector. Both the 80-kDa E1 protein and the 42.5-kDa E2 protein are nuclear phosphoproteins. The E1 and E2 proteins form a heteromeric complex within the insect cells, and both proteins localize to a DNA fragment which contains the viral origin of replication. In addition, we have detected an E1-associated ATPase and GTPase activity, which is likely part of an energy-generating system for the helicase activity which is predicted for this protein. The human papillomavirus type 11 E1 and E2 proteins possess the same replication-associated activities exhibited by the corresponding BPV proteins, suggesting that the replication activities of these viruses are tightly conserved.

MeSH Terms
Adenosine Triphosphatases/metabolism Amino Acid Sequence Animals Baculoviridae/genetics Base Sequence DNA Replication DNA, Viral/genetics Humans Macromolecular Substances Molecular Sequence Data Moths/cytology Oncogene Proteins, Viral/genetics,immunology,metabolism Papillomaviridae/genetics,metabolism Phosphoproteins/metabolism Phosphoric Monoester Hydrolases/metabolism Phosphorylation Plasmids/genetics Pyrophosphatases Recombinant Proteins/metabolism
Chemicals
DNA, Viral Macromolecular Substances Oncogene Proteins, Viral Phosphoproteins Recombinant Proteins Phosphoric Monoester Hydrolases Adenosine Triphosphatases Pyrophosphatases uridine triphosphatase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Bream G L
Division of Virology, Burroughs Wellcome Co., Research Triangle Park, North Carolina 27709.
Ohmstede C A
Phelps W C
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.67.5.2655-2663.1993
Published
1993-05-00
Pages
2655-63
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC237587
Subset
IM
External Links
PubMed source DOI: 10.1128/JVI.67.5.2655-2663.1993 PMC Full Text View journal details
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