1887

Abstract

GlnD is a pivotal protein in sensing intracellular levels of fixed nitrogen and has been best studied in enteric bacteria, where it reversibly uridylylates two related proteins, PII and GlnK. The uridylylation state of these proteins determines the activities of glutamine synthetase (GS) and NtrC. Results presented here demonstrate that is an essential gene in . Null mutations were introduced into the genome, but none could be stably maintained unless a second mutation was present that resulted in unregulated activity of GS. One mutation, , occurred spontaneously to give strain MV71, which failed to uridylylate the GlnK protein. The second, created by design, was (MV75), altering the adenylylation site of GS. The mutation is probably located in , encoding adenylyltransferase, because introducing the gene into MV72, a derivative of MV71, restored the regulation of GS activity. GlnK-UMP is therefore apparently required for GS to be sufficiently deadenylylated in for growth to occur. The Δ GS isolates were Nif, which could be corrected by introducing a mutation, confirming a role for GlnD in mediating gene regulation via some aspect of the NifL/NifA interaction. MV71 was unexpectedly NtrC, suggesting that NtrC activity might be regulated differently than in enteric organisms.

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2001-05-01
2019-10-19
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