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. 2020 Apr 7;48(9):5006–5015. doi: 10.1093/nar/gkaa205

Figure 3.

Figure 3.

AXP is required for the transition to DNA packaging. The DNA packaging reaction was performed in two steps as described in Materials and Methods and as outlined at top of the figure. (A) The cos-cleavage reaction (step 1) was performed in the absence of IHF but in the presence of the indicated nucleotide (50 μM). The first lane shows the products of the cos-cleavage reaction with the migration of un-cut substrate (S, 12.2 kb) and the two nuclease products (DL∼ 7.5 kb, DR∼ 4.7 kb) indicated. This was the input DNA for the packaging reaction (step 2) and DNase resistant (packaged) DNA is shown in the gel. Note that only the DL strand is packaged. (B) Same as Panel A except that IHF was included in step 1 of the reaction. The substrate was totally cut under these conditions (lane S). This was the input DNA for the packaging reaction (step 2) and DNase resistant (packaged) DNA is shown in the agarose gel. (C) Quantitation of the data presented in Panel B plus that for a packaging reaction containing 1 mM AMP-PCP in step 1. (D) The cos-cleavage reaction (step 1) was performed in the presence of IHF and in the presence of varying concentrations of ADP, as indicated, followed by DNA packaging (step 2). Each data point represents the average of three separate experiments with standard deviation shown in bars. The solid line represents the best fit of the data to a Langmuir binding model.