Lon-mediated DnaA proteolysis.

(A) DnaA in vivo stability assay of wild-type and lon::Ω cells in mid-exponential phase in M2G and while experiencing N or P starvation. (B) Quantification of DnaA in vivo stability assays done as shown in (A), from either five (M2G mid-exponential phase), two (N-limited), or three (P-limited) independent replicates for the lon::Ω strain, shown alongside wild-type data from Fig 3B, with error bars showing standard deviations. (C–E) Immunoblots of DnaA after shifting cells to nutrient-limited media, comparing lon::Ω to wild-type cells. For (C) and (D), the wild-type half of the blot was separated from the lon::Ω half, washed, and redeveloped to prevent the high signal of the lon::Ω bands from drowning out the wild-type signal. (F) DNA content as determined by flow cytometry of wild-type and lon::Ω cells after being shifted to C- or P-limited media. Each separate histogram represents 30 000 cells. (G) DNA content presented essentially as in (F), of wild-type and lon::Ω cells in mid-exponential phase in M2G before being shifted to N-limited medium (t = 0 h), and after being shifted to N-limited medium.

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