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In the cyanobacterium Synechococcus sp. strain PCC 7942 a multigene family of three different isozymes encodes the proteolytic subunit ClpP of the ATP-dependent Clp protease. In contrast to the monocistronic clpPI gene, clpPII and clpPIII are part of two bicistronic operons with clpX and clpR, respectively. Unlike most bacterial Clp proteins, the Synechococcus ClpP2, ClpP3, ClpR and ClpX proteins were not highly inducible by high temperatures, or by other stresses such as cold, high light or oxidation, although slower gradual rises occurred for all four proteins during high light, and for ClpP3, ClpR and ClpX at low temperature. Attempts to inactivate the clpPII, clpIII, clpR or clpX genes were only successful for clpPII, suggesting the others are essential for Synechococcus cell viability. The DeltaclpPII mutant exhibited no significant phenotypic changes from the wild-type, including no change in ClpX content. Despite the apparent bicistronic arrangement of both clpPII-clpX and clpR-clpPIII, all four genes primarily produce monocistronic transcripts, although polycistronic transcripts were detected. Mapping of 5' ends for the clpX and clpPIII monocistronic transcripts revealed promoters situated within the 3' region of clpPII and clpR, respectively. Transcriptional and translational studies further showed differences in the expression and regulation between the clpP-clpR-clpX genes. Inactivation of clpPI caused a significant decrease in ClpP2 protein concomitant to small increases in both ClpP3 and ClpR. Inactivation of clpPII resulted in a large rise in clpPI transcripts but to a lesser extent in ClpP1 protein. Similar small increases in ClpP3, ClpR and ClpX proteins also occurred in DeltaclpPII. These results highlight the regulatory complexity of these multiple clp genes and their functional importance in cyanobacteria.

microbiology

The clpP multigene family for the ATP-dependent Clp protease in the cyanobacterium Synechococcus c cThe GenBank accession numbers for the sequences of clpPII-clpX and clpR-clpPIII reported in this paper are U92039 and AJ132005, respectively.