Anabaena sp. strain PCC 7120 ntcA gene required for growth on nitrate and heterocyst development

Abstract

The Anabaena sp. strain PCC 7120 ntcA (bifA) gene encodes a sequence-specific DNA-binding protein, NtcA (BifA, VF1) that interacts with the upstream region of several genes, including glnA, xisA, rbcL, and nifH. We have constructed a ntcA null mutant by interrupting the gene with an omega Spr-Smr cassette. The ntcA mutant was not able to grow with nitrate or atmospheric dinitrogen as the sole nitrogen source but could be grown on medium containing ammonium. The ntcA mutant was unable to form heterocysts and did not rearrange the nifD or fdxN elements after induction on a medium lacking combined nitrogen. Northern (RNA) analysis of ntcA in the wild-type strain during nitrogen stepdown showed a peak of ntcA message at an early stage (12 h) of heterocyst induction. Complementation of the ntcA mutant with a DNA fragment containing the ntcA gene and 251 bp of upstream sequence on a shuttle vector restored a wild-type phenotype; however, a similar construction containing 87 bp of upstream sequence only partially restored the phenotype. Northern analysis of RNA samples isolated from ammonium-grown cultures of the ntcA mutant showed reduced amounts of glnA message and the absence of a 1.7-kb transcript. In the wild type, the 1.7-kb transcript represents the majority of glnA transcripts after nitrogen stepdown. The ntcA mutant showed a normal pattern of rbcLS messages under these growth conditions.