Uridylylation of the P(II) protein in the photosynthetic bacterium Rhodospirillum rubrum.
The regulatory protein P(II) has been studied in great detail in enteric bacteria; however, its function in photosynthetic bacteria has not been clearly established. As a number of these bacteria have been shown to regulate nitrogenase activity by a metabolic control system, it is of special interest to establish the role of P(II) in these diazotrophs. In this study, we show that P(II) in Rhodospirillum rubrum is modified in response to the N status in the cell and that addition of ammonium or glutamine leads to demodification. We also provide evidence that P(II) is uridylylated. In addition, we show that not only these compounds but also NAD+ promotes demodification of P(II), which is of particular interest as this pyridine nucleotide has been shown to act as a switch-off effector of nitrogenase. Demodification of P(II) by ammonium or NAD+ did not occur in cultures treated with an inhibitor of glutamine synthetase (methionine sulfoximine), whereas treatment with the glutamate synthase inhibitor 6-diazo-5-oxo-norleucine led to total demodification of P(II) without any other addition. The results indicate that P(II) probably is not directly involved in darkness switch-off of nitrogenase but that a role in ammonium switch-off cannot be excluded.
Johansson
M
M
Department of Biochemistry, Stockholm University, Sweden.
Nordlund
S
S
D001426:Bacterial Proteins / Q000378:metabolism*; D001483:Base Sequence; D001692:Biological Transport; D003980:Diazooxonorleucine / Q000494:pharmacology; D005974:Glutamate-Ammonia Ligase / Q000378:metabolism; D008717:Methionine Sulfoximine / Q000494:pharmacology; D008969:Molecular Sequence Data; D009584:Nitrogen / Q000378:metabolism; D009586:Nitrogen Fixation; D009713:Nucleotidyltransferases / Q000378:metabolism; D051922:PII Nitrogen Regulatory Proteins; D000644:Quaternary Ammonium Compounds; D012247:Rhodospirillum rubrum / Q000187:drug effects / Q000378:metabolism*; D014529:Uridine / Q000378:metabolism
D001426:Bacterial Proteins; D051922:PII Nitrogen Regulatory Proteins; D000644:Quaternary Ammonium Compounds; D003980:Diazooxonorleucine; D008717:Methionine Sulfoximine; C404143:PIID regulatory protein, Bacteria; D009713:Nucleotidyltransferases; C022515:regulatory protein uridylyltransferase; D005974:Glutamate-Ammonia Ligase; D009584:Nitrogen; D014529:Uridine