SeqA, the Escherichia coli origin sequestration protein, can regulate the replication of the pBR322 plasmid

• Published in: Plasmid Vol. 65; no. 1; pp. 15 - 19
• Main Authors: Douraid, Daghfous, Ahmed, Landoulsi
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 2011
• Subjects: adenine; Bacterial Outer Membrane Proteins - metabolism; DNA Methylation - genetics; DNA replication; DNA Replication - genetics; DNA, Bacterial - genetics; DNA-Binding Proteins - metabolism; Escherichia coli; Escherichia coli - genetics; Escherichia coli Proteins - metabolism; genes; Membrane; methylation; oriC; oripBR322; pBR322; plasmids; Plasmids - genetics; promoter regions; Promoter Regions, Genetic - genetics; replication origin; Replication Origin - genetics; RNA; RNA, Bacterial - genetics; SeqA; Sequestration; oriC; Membrane; SeqA; pBR322; oripBR322; Sequestration
• ISSN: 0147-619X; 1095-9890; 1095-9890
• DOI: 10.1016/j.plasmid.2010.09.003

The pBR322 plasmid origin replication and oriC show similar responses to adenine methylation. Both are subject to sequestration by membrane fractions. In fact, like the host origin oriC, the RNA II promoter region of pBR322 is regulated by methylation of three GATC adenine methylation sites. The SeqA gene product acts in the negative control of oriC by sequestration. We suggest that the role of SeqA protein in sequestration is similar to oriC region DNA. Hence, SeqA recognize the methylation state of the pBR322RNA II promoter region by direct DNA binding in vitro. Using the pOC42 plasmid, we show that SeqA binds exclusively to the hemimethylated form of the replication origin of the pBR322 plasmid. In addition, we suggested that the SeqA protein could modulate periodically the initiation of replication of the pBR322 plasmid. The later could be fixed by its origin sequence, on a hemimethylated state, during the initiation of the replication.