Formation of the preprimosome protects lambda O from RNA transcription-dependent proteolysis by ClpP/ClpX

Using the bacteriophage lambda DNA replication system, composed entirely of purified proteins, we have tested the accessibility of the short-lived lambda O protein to the ClpP/ClpX protease during the various stages of lambda DNA replication. We find that binding of lambda O protein to its orilambda DNA sequence, leading to the so-called "O-some" formation, largely inhibits its degradation. On the contrary, under conditions permissive for transcription, the lambda O protein bound to the orilambda sequence becomes largely accessible to ClpP/ClpX-mediated proteolysis. However, when the lambda O protein is part of the larger orilambda:O.P.DnaB preprimosomal complex, transcription does not significantly increase ClpP/ClpX-dependent lambda O degradation. These results show that transcription can stimulate proteolysis of a protein that is required for the initiation of DNA replication.