Effects of perturbations of the nitrogenase electron transfer chain on reversible ADP-ribosylation of nitrogenase Fe protein in Klebsiella pneumoniae strains bearing the Rhodospirillum rubrum dra operon.
The redox state of nitrogenase Fe protein is shown to affect regulation of ADP-ribosylation in Klebsiella pneumoniae strains transformed by plasmids carrying dra genes from Rhodospirillum rubrum. The dra operon encodes dinitrogenase reductase ADP-ribosyltransferase and dinitrogenase reductase-activating glycohydrolase, enzymes responsible for the reversible inactivation, via ADP-ribosylation, of nitrogenase Fe protein in R. rubrum. In bacteria containing the dra operon in their chromosomes, inactivation occurs in response to energy limitation or nitrogen sufficiency. The dra gene products, expressed at a low level in K. pneumoniae, enable transformants to reversibly ADP-ribosylate nitrogenase Fe protein in response to the presence of fixed nitrogen. The activities of both regulatory enzymes are regulated in vivo as described in R. rubrum. Genetic perturbations of the nitrogenase electron transport chain were found to affect the rate of inactivation of Fe protein. Strains lacking the electron donors to Fe protein (NifF or NifJ) were found to inactivate Fe protein more quickly than a strain with wild-type background. Deletion of nifD, which encodes a subunit of nitrogenase MoFe protein, was found to result in a slower inactivation response. No variation was found in the reactivation responses of these strains. It is concluded that the redox state of the Fe protein contributes to the regulation of the ADP-ribosylation of Fe protein.
Halbleib
C M
CM
Department of Biochemistry, University of Wisconsin-Madison, Madison, Wisconsin 53706, USA.
Zhang
Y
Y
Roberts
G P
GP
Ludden
P W
PW
D036002:ADP Ribose Transferases / Q000235:genetics / Q000378:metabolism*; D000246:Adenosine Diphosphate Ribose / Q000378:metabolism*; D004579:Electron Transport; D004789:Enzyme Activation; D015870:Gene Expression; D006026:Glycoside Hydrolases / Q000235:genetics / Q000378:metabolism*; D015151:Immunoblotting / Q000379:methods; D007658:Ketone Oxidoreductases / Q000235:genetics / Q000378:metabolism; D007711:Klebsiella pneumoniae / Q000235:genetics / Q000378:metabolism; D016296:Mutagenesis; D009699:N-Glycosyl Hydrolases; D009586:Nitrogen Fixation; D009591:Nitrogenase / Q000235:genetics / Q000378:metabolism*; D009876:Operon; D010088:Oxidoreductases / Q000378:metabolism*; D012247:Rhodospirillum rubrum / Q000201:enzymology* / Q000235:genetics
D000246:Adenosine Diphosphate Ribose; D010088:Oxidoreductases; D009591:Nitrogenase; C029130:nitrogenase reductase; D007658:Ketone Oxidoreductases; C039443:pyruvate-flavodoxin oxidoreductase; D036002:ADP Ribose Transferases; C057637:dinitrogenase reductase ADP-ribosyltransferase; D006026:Glycoside Hydrolases; D009699:N-Glycosyl Hydrolases; C036091:ADP-ribosyl-(dinitrogen reductase) hydrolase