Compartmentalization of phage ø29 DNA replication: Interaction between the primer terminal protein and the membrane-associated protein
| Autor: | Bravo, Alicia, Illana, Belén, Salas, Margarita |
|---|---|
| Přispěvatelé: | National Institutes of Health (US), Ministerio de Economía y Competitividad (España), European Commission, Fundación Ramón Areces |
| Rok vydání: | 2000 |
| Předmět: | |
| Zdroj: | Digital.CSIC. Repositorio Institucional del CSIC instname |
| Popis: | The bacteriophage 29 replication protein p1 (85 amino acids) is membrane associated in Bacillus subtilis-infected cells. The C-terminal 52 amino acid residues of p1 are sufficient for assembly into protofilament sheet structures. Using chemical cross-linking experiments, we demonstrate here that p1ΔC43, a C-terminally truncated p1 protein that neither associates with membranes in vivo nor self-interacts in vitro, can interact with the primer terminal protein (TP) in vitro. Like protein p1, plasmid-encoded protein p1ΔC43 reduces the rate of 29 DNA replication in vivo in a dosage-dependent manner. We also show that truncated p1 proteins that retain the N-terminal 42 amino acids, when present in excess, interfere with the in vitro formation of the TP⋅dAMP initiation complex in a reaction that depends on the efficient formation of a primer TP–29 DNA polymerase heterodimer. This interference is suppressed by increasing the concentration of either primer TP or 29 DNA polymerase. We propose a model for initiation of in vivo 29 DNA replication in which the viral replisome attaches to a membrane-associated p1-based structure. This work was supported by the National Institutes of Health (grant 5R01 GM27242-20), by the Dirección General de Investigación Científica y Técnica (grant PB98-0645) and by the European Union (BIO4-CT98-0250 and ERBFMX-CT97-0125). The institutional help of Fundación Ramón Areces is acknowledged. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|