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Structural and functional homology between the α and β subunits of the nitrogenase MoFe protein as revealed by sequencing the Rhizobium japonicum nifK gene

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Summary

Together with the nifK gene presented here all three genes encoding the polypeptides of the nitrogenase complex of Rhizobium japonicum are now sequenced. This allows a comprehensive comparison between them. The R. japonicum nifK gene has 518 codons predicting a MoFe protein β subunit with a calculated mol.wt. of 57428. Interspecies comparisons with one complete and two partial nifK sequences from other N2 fixing bacteria reveal conserved regions. The β subunit has three strongly conserved cysteine residues (at positions 69, 94, and 152). They are located in an area sharing amino acid sequence homology with the α subunit (the nifD gene product), which in turn has analogous cysteines at positions 67, 93, and 159. This may reflect the structural requirements of the α and β polypeptides for the binding of [4Fe:4S] clusters, and may indicate that nifD and nifK have evolved from a common ancestral gene. The comparative DNA sequence analysis also reveals structures required for nif gene expression in R. japonicum: (1) an apparent ribosome binding site with the consensus sequence 5′-uUgAaGGA-3′ is located six to eight nucleotides upstream of the AUG initiator codon; (2) nifK (being the promoter-distal gene of the nifDK operon) is followed by a potential ‘stem-and-loop’ transcriptional terminator.

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Authors and Affiliations

  1. Mikrobiologisches Institut, Eidgenössische Technische Hochschule, ETH-Zentrum, Universitätstrasse 2, CH-8092, Zürich, Switzerland

    Beat Thöny, Klaus Kaluza & Hauke Hennecke

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  1. Beat Thöny
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  2. Klaus Kaluza
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  3. Hauke Hennecke
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Communicated by H. Saedler

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Thöny, B., Kaluza, K. & Hennecke, H. Structural and functional homology between the α and β subunits of the nitrogenase MoFe protein as revealed by sequencing the Rhizobium japonicum nifK gene. Mol Gen Genet 198, 441–448 (1985). https://doi.org/10.1007/BF00332937

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  • DOI: https://doi.org/10.1007/BF00332937

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