scholarly journals Modulation of Bovine Papillomavirus DNA Replication by Phosphorylation of the Viral E1 Protein

Virology ◽  
1997 ◽  
Vol 228 (1) ◽  
pp. 1-10 ◽  
Author(s):  
THOMAS A ZANARDI ◽  
CHRISTINE M STANLEY ◽  
BRADLEY M SAVILLE ◽  
SUSAN M SPACEK ◽  
MICHAEL R LENTZ
Keyword(s):  
Dna Replication ◽  
Bovine Papillomavirus ◽  
E1 Protein ◽  
Papillomavirus Dna

scholarly journals Interactions of the Papovavirus DNA Replication Initiator Proteins, Bovine Papillomavirus Type 1 E1 and Simian Virus 40 Large T Antigen, with Human Replication Protein A

1999 ◽  
Vol 73 (6) ◽  
pp. 4899-4907 ◽  
Author(s):  
YuFeng Han ◽  
Yueh-Ming Loo ◽  
Kevin T. Militello ◽  
Thomas Melendy
Keyword(s):  
Dna Replication ◽  
Protein A ◽  
Simian Virus 40 ◽  
Simian Virus ◽  
T Antigen ◽  
Bovine Papillomavirus ◽  
Replication Proteins ◽  
E1 Protein ◽  
Cellular Dna

ABSTRACT Papovaviruses utilize predominantly cellular DNA replication proteins to replicate their own viral genomes. To appropriate the cellular DNA replication machinery, simian virus 40 (SV40) large T antigen (Tag) binds to three different cellular replication proteins, the DNA polymerase α-primase complex, the replication protein A (RPA) complex, and topoisomerase I. The functionally similar papillomavirus E1 protein has also been shown to bind to the DNA polymerase α-primase complex. Enzyme-linked immunoassay-based protein interaction assays and protein affinity pull-down assays were used to show that the papillomavirus E1 protein also binds to the cellular RPA complex in vitro. Furthermore, SV40 Tag was able to compete with bovine papillomavirus type 1 E1 for binding to RPA. Each of the three RPA subunits was individually overexpressed in Escherichia colias a soluble fusion protein. These fusion proteins were used to show that the E1-RPA and Tag-RPA interactions are primarily mediated through the 70-kDa subunit of RPA. These results suggest that different viruses have evolved similar mechanisms for taking control of the cellular DNA replication machinery.

scholarly journals Surface Mutagenesis of the Bovine Papillomavirus E1 DNA Binding Domain Reveals Residues Required for Multiple Functions Related to DNA Replication

2006 ◽  
Vol 80 (15) ◽  
pp. 7491-7499 ◽  
Author(s):  
Stephen Schuck ◽  
Arne Stenlund
Keyword(s):  
Dna Replication ◽  
Dna Binding ◽  
Binding Activity ◽  
Binding Domain ◽  
Bovine Papillomavirus ◽  
Dna Binding Domain ◽  
Multifunctional Protein ◽  
Dna Binding Activity ◽  
E1 Protein ◽  
Complex Functions

ABSTRACT The E1 protein from papillomaviruses is a multifunctional protein with complex functions required for the initiation of viral DNA replication. We have performed a surface mutagenesis of the well-characterized E1 DNA binding domain (DBD). We demonstrate that substitutions of multiple residues on the surface of the E1 DBD are defective for DNA replication without affecting the DNA binding activity of the protein. The defects of individual substitutions include failure to form the double trimer that melts the ori and failure to form the double hexamer that unwinds the ori. These results demonstrate that the DBD plays an essential role in multiple DNA replication-related processes apart from DNA binding.

scholarly journals Structure-Based Mutational Analysis of the Bovine Papillomavirus E1 Helicase Domain Identifies Residues Involved in the Nonspecific DNA Binding Activity Required for Double Trimer Formation

2010 ◽  
Vol 84 (9) ◽  
pp. 4264-4276 ◽  
Author(s):  
Xiaofei Liu ◽  
Stephen Schuck ◽  
Arne Stenlund
Keyword(s):  
Dna Replication ◽  
Dna Binding ◽  
Mutational Analysis ◽  
Binding Activity ◽  
Helicase Domain ◽  
Bovine Papillomavirus ◽  
Dna Binding Activity ◽  
E1 Protein ◽  
Initiation Of Dna Replication ◽  
Dna Template

ABSTRACT The papillomavirus E1 protein is a multifunctional initiator protein responsible for preparing the viral DNA template for initiation of DNA replication. The E1 protein encodes two DNA binding activities that are required for initiation of DNA replication. A well-characterized sequence-specific DNA binding activity resides in the E1 DBD and is used to tether E1 to the papillomavirus ori. A non-sequence-specific DNA binding activity is also required for formation of the E1 double trimer (DT) complex, which is responsible for the local template melting that precedes loading of the E1 helicase. This DNA binding activity is very poorly understood. We use a structure-based mutagenesis approach to identify residues in the E1 helicase domain that are required for the non-sequence-specific DNA binding and DT formation. We found that three groups of residues are involved in nonspecific DNA binding: the E1 β-hairpin structure containing R505, K506, and H507; a hydrophobic loop containing F464; and a charged loop containing K461 together generate the binding surface involved in nonspecific DNA binding. These residues are well conserved in the T antigens from the polyomaviruses, indicating that the polyomaviruses share this nonspecific DNA binding activity.

scholarly journals Bovine papillomavirus type 1 E2 protein heterodimer is functional in papillomavirus DNA replication in vivo

Virology ◽  
2009 ◽  
Vol 386 (2) ◽  
pp. 353-359 ◽  
Author(s):  
Reet Kurg ◽  
Piia Uusen ◽  
Toomas Sepp ◽  
Mari Sepp ◽  
Aare Abroi ◽  
...  
Keyword(s):  
Dna Replication ◽  
Bovine Papillomavirus ◽  
E2 Protein ◽  
Papillomavirus Dna

scholarly journals Induction of the Bovine Papillomavirus Origin “Onion Skin”-Type DNA Replication at High E1 Protein Concentrations In Vivo

2002 ◽  
Vol 76 (11) ◽  
pp. 5835-5845 ◽  
Author(s):  
Andres Männik ◽  
Kertu Rünkorg ◽  
Nele Jaanson ◽  
Mart Ustav ◽  
Ene Ustav
Keyword(s):  
Dna Replication ◽  
Skin Type ◽  
Bovine Papillomavirus ◽  
Two Dimensional Gel Electrophoresis ◽  
Reading Frame ◽  
E1 Protein ◽  
Onion Skin ◽  
Origin Region ◽  
Replication Intermediates

ABSTRACT We have studied the replication of plasmids composed of bovine papillomavirus type 1 (BPV1) origin of replication and expression cartridges for viral proteins E1 and E2 in hamster and mouse cells. We found that the replication mode changed dramatically at different expression levels of the E1 protein. At high levels of the E1 protein, overreplication of the origin region of the plasmid was observed. Analysis of the replication products by one-dimensional and two-dimensional gel electrophoresis suggested that initially “onion skin”-type replication intermediates were generated, presumably resulting from initiation of the new replication forks before the leading fork completed the synthesis of the DNA on the episomal plasmid. These replication intermediates served as templates for generation of a heterogeneous set of origin region-containing linear fragments by displacement synthesis at the partially replicated plasmid. Additionally, the linear fragments may have been generated by DNA break-up of the onion skin-type intermediates. Analysis of replication products indicated that generated linear fragments recombined and formed concatemers or circular molecules, which presumably were able to replicate in an E1- and E2-dependent fashion. At moderate and low levels of E1, generated by transcription of the E1 open reading frame using weaker promoters, DNA replication was initiated at much lower levels, which allowed elongation of the replication fork starting from the origin to be more balanced and resulted in the generation of full-sized replication products.

scholarly journals Nucleocytoplasmic Shuttling of Bovine Papillomavirus E1 Helicase Downregulates Viral DNA Replication in S Phase

2006 ◽  
Vol 81 (1) ◽  
pp. 384-394 ◽  
Author(s):  
Chiung-Yueh Hsu ◽  
Francisca Mechali ◽  
Catherine Bonne-Andrea
Keyword(s):  
Dna Replication ◽  
Viral Replication ◽  
Cyclin A ◽  
S Phase ◽  
Replication Capacity ◽  
Bovine Papillomavirus ◽  
Nucleocytoplasmic Shuttling ◽  
Viral Dna ◽  
Viral Dna Replication ◽  
E1 Protein

ABSTRACT The papillomavirus E1 protein is essential for the initiation of viral replication. We previously showed that the bovine papillomavirus E1 protein is unstable and becomes resistant to ubiquitin-mediated degradation when tightly bound to cyclin E-cyclin-dependent kinase 2 (Cdk2) before the start of DNA synthesis. However, neither the protection nor the targeted degradation of E1 appears to depend on its phosphorylation by Cdk. Here, we report that Cdk phosphorylation of E1 is also not a prerequisite for the initiation of viral DNA replication either in vitro or in vivo. Nevertheless, we found that phosphorylation of one Cdk site, Ser283, abrogates E1 replicative activity only in a cellular context. We show that this site-specific phosphorylation of E1 drives its export from the nucleus and promotes its continuous nucleocytoplasmic shuttling. In addition, we find that E1 shuttling occurs in S phase, when cyclin A-Cdk2 is activated. E1 interacts with the active cyclin A-Cdk2 complex and is phosphorylated on Ser283 by this kinase. These data suggest that the phosphorylation of E1 on Ser283 is a negative regulatory event that is involved in preventing the amplification of viral DNA during S phase. This finding reveals a novel facet of E1 regulation that could account for the variations of the viral replication capacity during different cell cycle phases, as well as in different stages of the viral cycle.

scholarly journals Transient Viral DNA Replication and Repression of Viral Transcription Are Supported by the C-Terminal Domain of the Bovine Papillomavirus Type 1 E1 Protein

1998 ◽  
Vol 72 (1) ◽  
pp. 796-801 ◽  
Author(s):  
Maureen C. Ferran ◽  
Alison A. McBride
Keyword(s):  
Dna Replication ◽  
Transcriptional Repression ◽  
Viral Transcription ◽  
Bovine Papillomavirus ◽  
Wild Type ◽  
Viral Dna ◽  
Viral Dna Replication ◽  
Terminal Domain ◽  
E1 Protein

ABSTRACT The bovine papillomavirus type 1 E1 protein is important for viral DNA replication and transcriptional repression. It has been proposed that the full-length E1 protein consists of a small N-terminal and a larger C-terminal domain. In this study, it is shown that an E1 polypeptide containing residues 132 to 605 (which represents the C-terminal domain) is able to support transient viral DNA replication, although at a level lower than that supported by the wild-type protein. This domain can also repress E2-mediated transactivation from the P89 promoter as well as the wild-type E1 protein can.

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