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J. Bacteriol. doi:10.1128/JB.01861-06
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Transcription of all amoC copies is associated with the recovery of Nitrosomonas europaea from ammonia starvation

Department of Microbiology, University of Washington; Department of Civil and Environmental Engineering, University of Washington

^* To whom correspondence should be addressed. Email: dastahl{at}u.washington.edu.

	Abstract

The chemolithotrophic ammonia-oxidizing bacterium, Nitrosomonas europaea, is known to be highly resistant to starvation conditions. The transcriptional response of N. europaea to ammonia addition following short and long-term starvation was examined by primer extension and S1 nuclease protection analyses of genes encoding enzymes for ammonia oxidation (amoCAB operons), CO₂ fixation (cbbLS), a third, lone copy of amoC (amoC₃), and two representative housekeeping genes (glyA and rpsJ). Primer extension analysis of RNA isolated from growing, starved, and recovering cells revealed two differentially regulated promoters upstream of the two amoCAB operons. The distal ⁷⁰ type amoCAB promoter was constitutively active in the presence of ammonia, but the proximal promoter was only active when cells were recovering from ammonia starvation. The lone, divergent copy of amoC (amoC₃) was expressed only during recovery. Both the proximal amoC_1,2 promoter and the amoC₃ promoter are similar to gram-negative ^E promoters, thus implicating ^E in the regulation of the recovery response. Although modeling of subunit interactions suggested that a non-conservative proline substitution in AmoC₃ may modify the activity of the holoenzyme, characterization of a amoC₃ strain showed no significant difference in starvation recovery under conditions evaluated. In contrast to the amo transcripts, a delayed appearance of transcripts for a gene required for CO₂ fixation (cbbL) suggested that its transcription is retarded until sufficient energy is available. Overall, these data revealed a programmed exit from starvation likely involving regulation by ^E and the coordinated regulation of catabolic and anabolic genes.

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