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. 2005 Mar 31;24(8):1502–1511. doi: 10.1038/sj.emboj.7600634

Figure 4.

Figure 4

A flexible linker between the N- and C-terminal domains enhances versatile DNA binding. (A) EMSAs of SeqA, SeqA-I21R, Δ1SeqA and Δ1SeqA-I21R with DNAs containing two hemimethylated GATC sequences. For each gel, the left-most lane contains an equimolar mixture of DNAs with 5, 7, 12, 21, 25 and 34 bp between the two GATC sites but no proteins. The following lanes show the interaction of various SeqA proteins with DNAs containing two GATC sequences separated by a variable number of bp (x), where x ranges from 4 to 34 bp. (B) Flow cytometry analysis reveals that the flexible linker between the two domains is partially dispensable for SeqA function in vivo. Basal expression of the plasmid encoding Δ1SeqA (pAG8020) restores replication synchrony of a ΔseqA∷tet strain. The plasmid encoding Δ1SeqA-I21R (pAG8025) with restricted flexibility between domains and unable to form filaments has asynchronous replication. Synchrony of the ΔseqA∷tet strain carrying the pAG8025 plasmid is restored by IPTG-induced protein overexpression. Arrowhead indicates a DNA content equivalent to two chromosomes.