Rapid assembly of the bacteriophage T4 core replication complex on a linear primer/template construct.
AUTOR(ES)
Kaboord, B F
RESUMO
DNA synthesis on a primed DNA substrate by bacteriophage T4 requires the assembly of a core replication complex consisting of the T4 DNA polymerase, a single-stranded binding protein (32 protein), and the accessory proteins 44/62 and 45. In this paper, we demonstrate the successful assembly of this core complex on a short linear primer/template system at levels of accessory proteins equivalent to the concentration of primer 3' ends. The key to this assembly is the presence of streptavidin molecules bound at each end of the DNA substrate via biotin moieties incorporated into the template strand. Streptavidin serves to block the ends of the primer/template, thus preventing translocation of the accessory proteins away from the site of assembly and their subsequent dissociation from the ends of the primer/template. Complex assembly on this substrate requires ATP and the presence of both the 44/62 and 45 proteins. The time required for assembly of a full enzyme equivalent of complex in our system is approximately 2 s.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=47882Documentos Relacionados
- Assembly of a functional replication complex without ATP hydrolysis: a direct interaction of bacteriophage T4 gp45 with T4 DNA polymerase.
- Prohead core of bacteriophage T4 can act as an intermediate in the T4 head assembly pathway.
- Mutational analysis of the tRNA3Lys/HIV-1 RNA (primer/template) complex.
- Early Intracellular Events in the Replication of Bacteriophage T4 Deoxyribonucleic Acid: V. Further Studies on the T4 Protein-Deoxyribonucleic Acid Complex 1
- Cyclin-Dependent Kinase Inhibitor p21 Modulates the DNA Primer-Template Recognition Complex