The right half of theEscherichia colireplication origin is not essential for viability, but facilitates multi-forked replication
| Autor: | Erik Boye, David Bates, Nicholas Stepankiw, Akihiro Kaidow |
|---|---|
| Rok vydání: | 2009 |
| Předmět: |
DNA Replication
DNA Bacterial Genetics Origin Recognition Complex DNA replication Replication Origin Biology Origin of replication Pre-replication complex Microbiology Article DnaA Licensing factor Plasmid SeqA protein domain Mutagenesis Escherichia coli bacteria Origin recognition complex Molecular Biology Sequence Deletion |
| Zdroj: | Molecular Microbiology. 74:467-479 |
| ISSN: | 1365-2958 0950-382X |
| DOI: | 10.1111/j.1365-2958.2009.06877.x |
| Popis: | Replication initiation is a key event in the cell cycle of all organisms and oriC, the replication origin in Escherichia coli, serves as the prototypical model for this process. The minimal sequence required for oriC function was originally determined entirely from plasmid studies using cloned origin fragments, which have previously been shown to differ dramatically in sequence requirement from the chromosome. Using an in vivo recombineering strategy to exchange wt oriCs for mutated ones regardless of whether they are functional origins or not, we have determined the minimal origin sequence that will support chromosome replication. Nearly the entire right half of oriC could be deleted without loss of origin function, demanding a reassessment of existing models for initiation. Cells carrying the new DnaA box-depleted 163 bp minimal oriC exhibited little or no loss of fitness under slow-growth conditions, but were sensitive to rich medium, suggesting that the dense packing of initiator binding sites that is a hallmark of prokaryotic origins, has likely evolved to support the increased demands of multi-forked replication. |
| Databáze: | OpenAIRE |
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