Table 3.
Quantification of the nucleotide dependency of the ParM mediated enhancement of pairing
| Proteins used | Nucleotide used | % of protein-bound DNA molecules present as
|
Molecules counted, n | |
|---|---|---|---|---|
| Pairs | Higher order complexes | |||
| ParR | — | 6.3 | 0.0 | 253 |
| ParR + ParM | — | 4.7 | 0.0 | 253 |
| ParR + ParM | ATP | 11 | 0.0 | 229 |
| ParR + ParM | ADP | 6.3 | 0.0 | 224 |
| ParR + ParM | AMP-PNP | 7.1 | 0.0 | 225 |
| ParR + ParM | ATPγS | 5.7 | 1.2 | 243 |
| ParR + ParM D170E | ATP | 5.6 | 0.0 | 259 |
ParR (100 ng), ParM (100 ng), and/or ParM D170E (100 ng) were bound to 100 ng linearized pMD330 DNA in the presence of the indicated nucleotides and prepared for electron microscopy. AMP-PNP and ATPγS are nonhydrolysable ATP analogs. The frequencies of protein-bound DNA molecules present in paired structures or higher-order-complexes were determined by counting randomly selected fields. Shown are the results from a single, representative experiment. AMP-PNP, adenylyl-imidophosphate; ATPγS, adenosine-5′-O-thiotriphosphate.