Expression and functional analysis of an N‐truncated NifA protein of Herbaspirillum seropedicae
Open Access
- 26 March 1999
- journal article
- Published by Wiley in FEBS Letters
- Vol. 447 (2-3), 283-286
- https://doi.org/10.1016/s0014-5793(99)00314-2
Abstract
In Herbaspirillum seropedicae, an endophytic diazotroph, nif gene expression is under the control of the transcriptional activator NifA. We have over‐expressed and purified a protein containing the central and C‐terminal domains of the H. seropedicae NifA protein, N‐truncated NifA, fused to a His‐Tag sequence. This fusion protein was found to be partially soluble and was purified by affinity chromatography. Band shift and footprinting assays showed that the N‐truncated NifA protein was able to bind specifically to the H. seropedicae nifB promoter region. In vivo analysis showed that this protein activated the nifH promoter of Klebsiella pneumoniae in Escherichia coli only in the absence of oxygen and this activation was not negatively controlled by ammonium ions.Keywords
This publication has 21 references indexed in Scilit:
- Purification and binding analysis of the nitrogen fixation regulatory NifA protein from Azospirillum brasilenseBrazilian Journal of Medical and Biological Research, 1998
- The oxygen-responsive NIFL-NIFA complex: a novel two-component regulatory system controlling nitrogenase synthesis in γ-ProteobacteriaArchiv für Mikrobiologie, 1998
- DNA‐binding domain mutants of sigma‐N (σ N , σ 54 ) defective between closed and stable open promoter complex formationMolecular Microbiology, 1997
- Sequence and structural organization of a nifA-like gene and part of a nifB-like gene of Herbaspirillum seropedicae strain Z78Journal of General Microbiology, 1991
- The role of activator binding sites in transcriptional control of the divergently transcribed nifF and nif LA promoters from Klebsiella pneumoniaeMolecular Microbiology, 1988
- Characterization of Herbaspirillum seropedicae gen. nov., sp. nov., a Root-Associated Nitrogen-Fixing BacteriumInternational Journal of Systematic and Evolutionary Microbiology, 1986
- A Rhizobium meliloti symbiotic regulatory geneCell, 1984
- A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye bindingAnalytical Biochemistry, 1976
- Derivation and Properties of F-prime Factors in Escherichia coli Carrying Nitrogen Fixation Genes from Klebsiella pneumoniaeJournal of General Microbiology, 1976
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970