Maximal unwinding activity is approximately 19% for this substrate, suggesting that the partial duplex DNA lacks structural elements required for efficient PriA binding and unwinding (Figure 3). This has been observed for E. coli PriA helicase as well [7, 28]. Overall, these results demonstrate

that N. gonorrhoeae PriA helicase activity is limited to relatively short stretches of duplex DNA, akin to its E. coli counterpart. Figure 3 Helicase activity of N. gonorrhoeae PriA. PriA-catalyzed duplex DNA unwinding was examined using 1 nM Fork 1 (15 bp lagging strand arm, diamonds), Fork 2 (25 bp lagging strand arm, triangles), Fork 3 (40 bp lagging strand arm, squares), or 3′ Overhang BAY 73-4506 purchase (25 bp partial duplex, circles). Measurements are reported in triplicate

and error bars represent one standard deviation of the mean. Comparison of the helicase activity of N. gonorrhoeae PriA that we measured in this study with the previously reported helicase activity of E. coli PriA at the same concentrations and on similar DNA substrates reveals that the two PriA homologs follow the same trend with respect to the dependence of their DNA unwinding activity on the length of the duplex arm of the DNA GSK1210151A mw Substrate (Table 3). There are some differences in the degree of DNA unwinding catalyzed by N. gonorrhoeae PriA that we measured in this study compared with the helicase activity previously reported for E. coli PriA. For example, E. coli PriA helicase shows slightly elevated DNA unwinding activity on the 25 bp fork structure compared to N. gonorrhoeae PriA {Selleck Anti-diabetic Compound Library|Selleck Antidiabetic Compound Library|Selleck Anti-diabetic Compound Library|Selleck Antidiabetic Compound Library|Selleckchem Anti-diabetic Compound Library|Selleckchem Antidiabetic Compound Library|Selleckchem Anti-diabetic Compound Library|Selleckchem Antidiabetic Compound Library|Anti-diabetic Compound Library|Antidiabetic Compound Library|Anti-diabetic Compound Library|Antidiabetic Compound Library|Anti-diabetic Compound Library|Antidiabetic Compound Library|Anti-diabetic Compound Library|Antidiabetic Compound Library|Anti-diabetic Compound Library|Antidiabetic Compound Library|Anti-diabetic Compound Library|Antidiabetic Compound Library|Anti-diabetic Compound Library|Antidiabetic Compound Library|Anti-diabetic Compound Library|Antidiabetic Compound Library|Anti-diabetic Compound Library|Antidiabetic Compound Library|buy Anti-diabetic Compound Library|Anti-diabetic Compound Library ic50|Anti-diabetic Compound Library price|Anti-diabetic Compound Library cost|Anti-diabetic Compound Library solubility dmso|Anti-diabetic Compound Library purchase|Anti-diabetic Compound Library manufacturer|Anti-diabetic Compound Library research buy|Anti-diabetic Compound Library order|Anti-diabetic Compound Library mouse|Anti-diabetic Compound Library chemical structure|Anti-diabetic Compound Library mw|Anti-diabetic Compound Library molecular weight|Anti-diabetic Compound Library datasheet|Anti-diabetic Compound Library supplier|Anti-diabetic Compound Library in vitro|Anti-diabetic Compound Library cell line|Anti-diabetic Compound Library concentration|Anti-diabetic Compound Library nmr|Anti-diabetic Compound Library in vivo|Anti-diabetic Compound Library clinical trial|Anti-diabetic Compound Library cell assay|Anti-diabetic Compound Library screening|Anti-diabetic Compound Library high throughput|buy Antidiabetic Compound Library|Antidiabetic Compound Library ic50|Antidiabetic Compound Library price|Antidiabetic Compound Library cost|Antidiabetic Compound Library solubility dmso|Antidiabetic Compound Library purchase|Antidiabetic Compound Library manufacturer|Antidiabetic Compound Library research buy|Antidiabetic Compound Library order|Antidiabetic Compound Library chemical structure|Antidiabetic Compound Library datasheet|Antidiabetic Compound Library supplier|Antidiabetic Compound Library in vitro|Antidiabetic Compound Library cell line|Antidiabetic Compound Library concentration|Antidiabetic Compound Library clinical trial|Antidiabetic Compound Library cell assay|Antidiabetic Compound Library screening|Antidiabetic Compound Library high throughput|Anti-diabetic Compound high throughput screening| (Table 3). Whether this represents natural biological variation between the two PriA homologs or differences arising from work involving separate investigators is uncertain. Table 3 Comparison of helicase activity of E. coli PriA and N. gonorrhoeae PriA. DNA Substrate E. coli PriA1 % DNA

Unwound N. gonorrhoeae Diflunisal PriA2 % DNA Unwound 25 bp fork 83 ± 3 61 ± 6 40 bp fork 28 ± 8 37 ± 7 25 bp partial duplex 23 ± 2 17 ± 4 1Cadman et al. J Biol Chem 2005, 280(48):39693-39700. 2This study. In this study, the 25 bp fork substrate is Fork 2, the 40 bp fork substrate is Fork 3, and the 25 bp partial duplex substrate is 3′ Overhang. The helicase activity for each PriA homolog is the mean percent of DNA unwound by 5 nM PriA on 1 nM DNA substrate and in the absence of its cognate PriB. Mean values from Cadman et al. are derived from two independent experiments, and mean values from this study are derived from three independent experiments. Associated uncertainty values are one standard deviation of the mean. PriB stimulates PriA’s helicase activity on long regions of duplex DNA To determine if N. gonorrhoeae PriB stimulates the helicase activity of its cognate PriA, we examined PriA helicase activity on a forked DNA substrate with a 40 bp lagging strand arm (Fork 3) in the presence and absence of PriB.