Two cellular single-strand-specific DNA-binding proteins interact with two regions of the bovine papillomavirus type 1 genome, including the origin of DNA replication.

1992 ◽  
Vol 66 (10) ◽  
pp. 5988-5998 ◽  
Author(s):  
C Habiger ◽  
G Stelzer ◽  
U Schwarz ◽  
E L Winnacker
Keyword(s):  
Dna Replication ◽  
Dna Binding ◽  
Binding Proteins ◽  
Dna Binding Proteins ◽  
Single Strand ◽  
Bovine Papillomavirus ◽  
Origin Of Dna Replication

scholarly journals Sequence-specific interactions between cellular DNA-binding proteins and the adenovirus origin of DNA replication.

1987 ◽  
Vol 7 (2) ◽  
pp. 875-886 ◽  
Author(s):  
P J Rosenfeld ◽  
E A O'Neill ◽  
R J Wides ◽  
T J Kelly
Keyword(s):  
Dna Replication ◽  
Dna Binding ◽  
Affinity Chromatography ◽  
Binding Proteins ◽  
Dna Recognition ◽  
Dna Binding Proteins ◽  
Cellular Protein ◽  
Recognition Site ◽  
Base Substitution ◽  
Origin Of Dna Replication

The adenovirus origin of DNA replication contains three functionally distinct sequence domains (A, B, and C) that are essential for initiation of DNA synthesis. Previous studies have shown that domain B contains the recognition site for nuclear factor I (NF-I), a cellular protein that is required for optimal initiation. In the studies reported here, we used highly purified NF-I, prepared by DNA recognition site affinity chromatography (P. J. Rosenfeld and T. J. Kelly, Jr., J. Biol. Chem. 261:1398-1408, 1986), to investigate the cellular protein requirements for initiation of viral DNA replication. Our data demonstrate that while NF-I is essential for efficient initiation in vitro, other cellular factors are required as well. A fraction derived from HeLa cell nuclear extract (BR-FT fraction) was shown to contain all the additional cellular proteins required for the complete reconstitution of the initiation reaction. Analysis of this complementing fraction by a gel electrophoresis DNA-binding assay revealed the presence of two site-specific DNA-binding proteins, ORP-A and ORP-C, that recognized sequences in domains A and C, respectively, of the viral origin. Both proteins were purified by DNA recognition site affinity chromatography, and the boundaries of their binding sites were defined by DNase I footprint analysis. Additional characterization of the recognition sequences of ORP-A, NF-I, and ORP-C was accomplished by determining the affinity of the proteins for viral origins containing deletion and base substitution mutations. ORP-C recognized a sequence between nucleotides 41 and 51 of the adenovirus genome, and analysis of mutant origins indicated that efficient initiation of replication is dependent on the presence of a high-affinity ORP-C-binding site. The ORP-A recognition site was localized to the first 12 base pairs of the viral genome within the minimal origin of replication. These data provide evidence that the initiation of adenovirus DNA replication involves multiple protein-DNA interactions at the origin.

Sequence-specific interactions between cellular DNA-binding proteins and the adenovirus origin of DNA replication

1987 ◽  
Vol 7 (2) ◽  
pp. 875-886
Author(s):  
P J Rosenfeld ◽  
E A O'Neill ◽  
R J Wides ◽  
T J Kelly
Keyword(s):  
Dna Replication ◽  
Dna Binding ◽  
Affinity Chromatography ◽  
Binding Proteins ◽  
Dna Recognition ◽  
Dna Binding Proteins ◽  
Cellular Protein ◽  
Recognition Site ◽  
Base Substitution ◽  
Origin Of Dna Replication

The adenovirus origin of DNA replication contains three functionally distinct sequence domains (A, B, and C) that are essential for initiation of DNA synthesis. Previous studies have shown that domain B contains the recognition site for nuclear factor I (NF-I), a cellular protein that is required for optimal initiation. In the studies reported here, we used highly purified NF-I, prepared by DNA recognition site affinity chromatography (P. J. Rosenfeld and T. J. Kelly, Jr., J. Biol. Chem. 261:1398-1408, 1986), to investigate the cellular protein requirements for initiation of viral DNA replication. Our data demonstrate that while NF-I is essential for efficient initiation in vitro, other cellular factors are required as well. A fraction derived from HeLa cell nuclear extract (BR-FT fraction) was shown to contain all the additional cellular proteins required for the complete reconstitution of the initiation reaction. Analysis of this complementing fraction by a gel electrophoresis DNA-binding assay revealed the presence of two site-specific DNA-binding proteins, ORP-A and ORP-C, that recognized sequences in domains A and C, respectively, of the viral origin. Both proteins were purified by DNA recognition site affinity chromatography, and the boundaries of their binding sites were defined by DNase I footprint analysis. Additional characterization of the recognition sequences of ORP-A, NF-I, and ORP-C was accomplished by determining the affinity of the proteins for viral origins containing deletion and base substitution mutations. ORP-C recognized a sequence between nucleotides 41 and 51 of the adenovirus genome, and analysis of mutant origins indicated that efficient initiation of replication is dependent on the presence of a high-affinity ORP-C-binding site. The ORP-A recognition site was localized to the first 12 base pairs of the viral genome within the minimal origin of replication. These data provide evidence that the initiation of adenovirus DNA replication involves multiple protein-DNA interactions at the origin.

scholarly journals DNA-Binding Proteins Essential for Protein-Primed Bacteriophage Φ29 DNA Replication

2016 ◽  
Vol 3 ◽  
Author(s):  
Margarita Salas ◽  
Isabel Holguera ◽  
Modesto Redrejo-Rodríguez ◽  
Miguel de Vega
Keyword(s):  
Dna Replication ◽  
Dna Binding ◽  
Binding Proteins ◽  
Dna Binding Proteins

scholarly journals Single strand DNA binding proteins 1 and 2 protect newly replicated telomeres

Cell Research ◽  
2013 ◽  
Vol 23 (5) ◽  
pp. 705-719 ◽  
Author(s):  
Peili Gu ◽  
Wei Deng ◽  
Ming Lei ◽  
Sandy Chang
Keyword(s):  
Dna Binding ◽  
Binding Proteins ◽  
Dna Binding Proteins ◽  
Single Strand

scholarly journals Competition for DNA Binding Sites between the Short and Long Forms of E2 Dimers Underlies Repression in Bovine Papillomavirus Type 1 DNA Replication Control

1998 ◽  
Vol 72 (3) ◽  
pp. 1931-1940 ◽  
Author(s):  
Daniel A. Lim ◽  
Manfred Gossen ◽  
Chris W. Lehman ◽  
Michael R. Botchan
Keyword(s):  
Dna Replication ◽  
Dna Binding ◽  
Short Form ◽  
Bovine Papillomavirus ◽  
Binding Studies ◽  
Proliferating Cells ◽  
Dna Binding Sites ◽  
Replication Control

ABSTRACT Papillomaviruses establish a long-term latency in vivo by maintaining their genomes as nuclear plasmids in proliferating cells. Bovine papillomavirus type 1 encodes two proteins required for viral DNA replication: the helicase E1 and the positive regulator E2. The homodimeric E2 is known to cooperatively bind to DNA with E1 to form a preinitiation complex at the origin of DNA replication. The virus also codes for two short forms of E2 that can repress viral functions when overexpressed, and at least one copy of the repressor is required for stable plasmid maintenance in transformed cells. Employing a tetracycline-regulated system to control E1 and E2 production from integrated loci, we show that the short form of E2 negatively regulates DNA replication. We also found that the short form could repress replication in a cell-free replication system and that the repression requires the DNA binding domain of the protein. In contrast, heterodimers of the short and long forms were activators and, by footprint analysis, were shown to be as potent as homodimeric E2 in loading E1 to its cognate site. DNA binding studies show that when E1 levels are low and are dependent upon E2 for occupancy of the origin site, the repressor can block E1-DNA interactions. We conclude that DNA replication modulation results from competition between the different forms of E2 for DNA binding. Given that heterodimers are active and that the repressor form of E2 shows little cooperativity with E1 for DNA binding, this protein is a weak repressor.

Sign in / Sign up

Export Citation Format

Share Document