Characterization of human papillomavirus type 16 E2 protein and subdomains expressed in insect cells.

2.50
Hdl Handle:
http://hdl.handle.net/10541/97966
Title:
Characterization of human papillomavirus type 16 E2 protein and subdomains expressed in insect cells.
Authors:
Sanders, C M; Stern, Peter L; Maitland, N J
Abstract:
The E2 open reading frame of human papillomavirus type 16 (HPV-16) encodes a DNA-binding protein which modulates papillomavirus transcription and replication. To investigate the biological and biochemical properties of the HPV-16 E2 protein, we have constructed recombinant baculoviruses which express the full-length molecule and individual N- and C-terminal domains in Sf21 insect cells. In this system the full-length E2 protein was phosphorylated and targeted to the insect cell nucleus. A 93 amino acid C-terminal fragment encompassing the DNA binding and dimerization functions of E2 was also translocated to the nucleus but was not modified by phosphorylation. The E2 N-terminal protein accumulated in the insect cell cytoplasm but was not efficiently phosphorylated. The formation of heterodimers between full-length and N-terminally truncated E2 species was observed when Sf21 cells were co-infected with recombinant viruses and when homodimers were mixed in vitro, suggesting that the dimer interface is not sufficiently stable to prevent subunit exchange in vivo. Both homo- and heterodimeric E2 species were able to bind specifically and in any combination to tandem E2 binding sites from the HPV-16 regulatory region. Furthermore, the HPV-16 E2 protein bound to DNA exhibited a distinct susceptibility profile to pronase digestion, potentially contrasting with that reported for BPV-1 E2. These observations suggest that significant structural and functional differences may exist between the BPV/HPV E2 proteins and have implications for understanding E2-dependent regulation of transcription and replication.
Affiliation:
Department of Biology, University of York, Heslington, United Kingdom.
Citation:
Characterization of human papillomavirus type 16 E2 protein and subdomains expressed in insect cells. 1995, 211 (2):418-33 Virology
Journal:
Virology
Issue Date:
20-Aug-1995
URI:
http://hdl.handle.net/10541/97966
DOI:
10.1006/viro.1995.1424
PubMed ID:
7645246
Type:
Article
Language:
en
ISSN:
0042-6822
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorSanders, C Men
dc.contributor.authorStern, Peter Len
dc.contributor.authorMaitland, N Jen
dc.date.accessioned2010-05-05T10:53:11Z-
dc.date.available2010-05-05T10:53:11Z-
dc.date.issued1995-08-20-
dc.identifier.citationCharacterization of human papillomavirus type 16 E2 protein and subdomains expressed in insect cells. 1995, 211 (2):418-33 Virologyen
dc.identifier.issn0042-6822-
dc.identifier.pmid7645246-
dc.identifier.doi10.1006/viro.1995.1424-
dc.identifier.urihttp://hdl.handle.net/10541/97966-
dc.description.abstractThe E2 open reading frame of human papillomavirus type 16 (HPV-16) encodes a DNA-binding protein which modulates papillomavirus transcription and replication. To investigate the biological and biochemical properties of the HPV-16 E2 protein, we have constructed recombinant baculoviruses which express the full-length molecule and individual N- and C-terminal domains in Sf21 insect cells. In this system the full-length E2 protein was phosphorylated and targeted to the insect cell nucleus. A 93 amino acid C-terminal fragment encompassing the DNA binding and dimerization functions of E2 was also translocated to the nucleus but was not modified by phosphorylation. The E2 N-terminal protein accumulated in the insect cell cytoplasm but was not efficiently phosphorylated. The formation of heterodimers between full-length and N-terminally truncated E2 species was observed when Sf21 cells were co-infected with recombinant viruses and when homodimers were mixed in vitro, suggesting that the dimer interface is not sufficiently stable to prevent subunit exchange in vivo. Both homo- and heterodimeric E2 species were able to bind specifically and in any combination to tandem E2 binding sites from the HPV-16 regulatory region. Furthermore, the HPV-16 E2 protein bound to DNA exhibited a distinct susceptibility profile to pronase digestion, potentially contrasting with that reported for BPV-1 E2. These observations suggest that significant structural and functional differences may exist between the BPV/HPV E2 proteins and have implications for understanding E2-dependent regulation of transcription and replication.en
dc.language.isoenen
dc.subject.meshAmino Acid Sequence-
dc.subject.meshAnimals-
dc.subject.meshBase Sequence-
dc.subject.meshCell Line-
dc.subject.meshCloning, Molecular-
dc.subject.meshDNA Primers-
dc.subject.meshDNA-Binding Proteins-
dc.subject.meshEndopeptidases-
dc.subject.meshEnhancer Elements, Genetic-
dc.subject.meshMolecular Sequence Data-
dc.subject.meshNucleopolyhedrovirus-
dc.subject.meshOncogene Proteins, Viral-
dc.subject.meshPapillomaviridae-
dc.subject.meshPhosphorylation-
dc.subject.meshRecombinant Proteins-
dc.subject.meshSpodoptera-
dc.titleCharacterization of human papillomavirus type 16 E2 protein and subdomains expressed in insect cells.en
dc.typeArticleen
dc.contributor.departmentDepartment of Biology, University of York, Heslington, United Kingdom.en
dc.identifier.journalVirologyen
All Items in Christie are protected by copyright, with all rights reserved, unless otherwise indicated.