FIG 6.

Characterization of proteins associated with the C. crescentus RNA degradosome at low temperature. (A) Cultures of C. crescentus were grown at either 30°C or 15°C up to mid-log phase, and RNA degradosomes were isolated. The strains used were NA1000 containing FLAG-tagged RNase E (FLAG-RNE) and NA1000 with no tagged proteins (wt) as a negative control. The proteins indicated by arrowheads were identified by mass spectrometry. (B) Association of Rho with the C. crescentus RNA degradosome. Cultures of C. crescentus were grown at either 30°C (cold−) or 15°C (cold+) up to mid-log phase, and proteins from total cell extracts or isolated RNA degradosomes were separated by SDS-PAGE. The proteins were transferred to a PVDF membrane, and the Rho protein was identified by immunoblotting with an anti-Rho serum. The samples used were total extract of NA1000 (wt) and isolated RNA degradosomes from NA1000 (wt) or MM71 (ΔrhlB) containing FLAG-tagged RNase E (FLAG-RNE).