The P1 ParA protein and its ATPase activity play a direct role in the segregation of plasmid copies to daughter cells.
The P1 ParA protein is an ATPase that recognizes the parA promoter region where it acts to autoregulate the P1 parA-parB operon. The ParB protein is essential for plasmid partition and recognizes the cis-acting partition site parS. The regulatory role of ParA is also essential because a controlled level of ParB protein is critical for partition. However, we show that this regulatory activity is not the only role for ParA in partition. Efficient partition can be achieved without autoregulation as long as Par protein levels are kept within a range of low values. The properties of ParA mutants in these conditions showed that ParA is essential for some critical step in the partition process that is independent of par operon regulation. The putative nucleotide-binding site for the ParA ATPase was identified and disrupted by mutation. The resulting mutant was substantially defective for autoregulation and completely inactive for partition in a system in which the need for autoregulation is abolished. Thus, the ParA nucleotide-binding site appears to be necessary both for the repressor activity of ParA and for some essential step in the partition process itself. We propose that the nucleotide-bound form of the enzyme adopts a configuration that favours binding to the operator, but that the ATPase activity of ParA is required for some energetic step in partition of the plasmid copies to daughter cells.
Davis
M A
MA
ABL-Basic Research Program, NCI-Frederick Cancer Research and Development Centre, Maryland 21702-1201, USA.
Radnedge
L
L
Martin
K A
KA
Hayes
F
F
Youngren
B
B
Austin
S J
SJ
D000251:Adenosine Triphosphatases / Q000235:genetics* / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D018983:DNA Footprinting; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development; D015964:Gene Expression Regulation, Bacterial; D008969:Molecular Sequence Data; D009154:Mutation; D009875:Operator Regions, Genetic; D009876:Operon; D010957:Plasmids / Q000235:genetics*; D011401:Promoter Regions, Genetic; D011485:Protein Binding
D001426:Bacterial Proteins; C052883:chromosome partition proteins, bacterial; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases