Abstract
Glutamine synthetase (GS; EC 6.3.1.2) activity from the unicellular cyanobacterium Synechococcus sp. strain PCC 6301 shows a short-term regulation by light-dark transitions. The enzyme activity declines down to 30% of the original level after 2 h of dark incubation, and can be fully reactivated within 15 min of re-illumination. The loss of activity is not due to protein degradation, but rather to a reversible change of the enzyme, as deduced from the GS-protein levels determined in dark-incubated cells using polyclonal antibodies raised against Synechococcus GS. Incubation with 3-(3-4-dichlorophenyl)-1,1-dimethylurea (DCMU) also provokes GS inactivation, indicating that an active electron flow between both photosystems is necessary to maintain GS in an active state. On the other hand, the light-mediated reactivation of GS in dark-incubated cells treated with dicyclohexyl-carbodiimide (DCCD) or carbonyl cyanide m-chlorophenylhydrazone (CCCP) indicates that neither changes in the ATP synthesis nor the lack of an electrochemical proton gradient across the thylakoid membrane are directly involved in the regulation process. The inactive form of GS is extremely labile in vitro after disruption of the cells, and is not reactivated by treatment with dithiothreitol or spinach thioredoxin m. These results, taken together with the fact that dark-promoted GS inactivation is dependent on the growth phase, seem to indicate that GS activity is not regulated by a typical redox process and that some other metabolic signal(s), probably related to the ammonium-assimilation pathway, might be involved in the regulation process. In this regard, our results indicate that glutamine is not a regulatory metabolite of Synechococcus glutamine synthetase.
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Abbreviations
- CAP:
-
chloramphenicol
- CCCP:
-
carbonyl cyanide m-chlorophenylhydrazone
- DCCD:
-
dicyclohexylcarbodiimide
- DCMU:
-
3-(3-4-dichlorophenyl)-1,1-dimethylurea
- DTT:
-
dithiothreitol
- GOGAT:
-
glutamate synthase
- GS:
-
glutamine synthetase
- PFD:
-
photon flux density
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This work has been financed by the Directión General de Investigación Científica y Técnica, (Grant PB88-0020) and by the Junta de Andalucía, Spain.
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Marqués, S., Mérida, A., Candau, P. et al. Light-mediated regulation of glutamine synthetase activity in the unicellular cyanobacterium Synechococcus sp. PCC 6301. Planta 187, 247–253 (1992). https://doi.org/10.1007/BF00201947
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DOI: https://doi.org/10.1007/BF00201947