Alternative Nitrogenases in Anabaena variabilis: The Role of Molybdate and Vanadate in Nitrogenase Gene

Abstract

Anabaena variabilis ATCC 29413 has two distinct nitrogenases that function in heterocysts, a conventional Mo-nitrogenase and an alternative V-nitrogenase. Synthesis of these two enzymes was repressed in cells growing with a source of fixed nitrogen, such as ammonium; however, the V-nitrogenase was also repressed by Mo. Expression of the V-nitrogenase which was not affected by V and expression of the Mo-nitrogenase was not affected by the presence or absence of either Mo or V. In the absence of both Mo and V in an environment lacking fixed nitrogen, cells became starved for both metals; however, low levels of nitrogen fixation and slow growth persisted. A mutant lacking the V-nitrogenase was still able to grow very slowly in Mo-and V-free medium; however, loss of the Mo-nitrogenase in a nifDK1 mutant abolished the residual growth, suggesting that only the Mo-nitrogenase functioned under these conditions to support slow growth. The addition of vanadate, molybdate, or tungstate, which is transported by the molybdate transporter, to cells starved for these metals resulted in an increase in nitrogenase activity within two hours after the addition of the metal and this increase required new protein synthesis. While tungstate functioned about as well as vanadate in supporting acetylene reduction, the cells were not able to grow any better with tungstate than with no added metal. A mutant lacking the V-nitrogenase showed no increase in nitrogenase activity upon addition of tungstate, suggesting that the V-nitrogenase was able to incorporate tungstate. Tungstate was able to substitute for molybdate in repressing transcription of a Mo-transport gene, but it did not repress transcription of the vnfH gene, which was repressed by Mo. The availability of Mo and V plays an important role in controlling whether the Mo-or the V-nitrogenase is used for nitrogen fixation.

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T. Thiel and B. Pratte, "Alternative Nitrogenases in Anabaena variabilis: The Role of Molybdate and Vanadate in Nitrogenase Gene," Advances in Microbiology, Vol. 3 No. 6A, 2013, pp. 87-95. doi: 10.4236/aim.2013.36A011.

Conflicts of Interest

The authors declare no conflicts of interest.

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