ClpS modulates but is not essential for bacterial N-end rule degradation

In eubacteria, the ClpS adaptor has been proposed to be essential for degradation of N-end rule substrates by the AAA(+) protease ClpAP. To test this model, we assayed degradation of substrates bearing N-end rule sequences isolated in a genetic screen for efficient degradation tags. ClpS was not vital for degradation in vivo but rather stimulated turnover in a sequence-specific manner. Although ClpS substantially enhanced degradation of N-end substrates at low substrate concentrations in vitro, it suppressed the degradation rate when substrate was saturating. Thus, we conclude that ClpAP recognizes N-end rule substrates directly, whereas ClpS modulates this degradation pathway.