Abstract
An E. coli groEL mutant transformed with the Klebsiella pneumoniae nif gene cluster accumulated very low to non-detectable levels of Kp1 and Kp2 as compared to the isogenic wild type strain. In K. pneumoniae, overexpression of the E. coli groE operon markedly accelerated the rate of appearance of Kpl and its constituent polypeptides after the start of derepression. NifA-dependent expression of β-galactosidase from the nifH promoter followed by an in-frame nifH‘-’lacZ fusion was threefold lower in the groEL mutant compared to the wild type strain. A direct interaction between NifA and GroEL was indicated by co-immunoprecipitation of NifA with anti-GroEL antibodies. Pulse-chase analysis of derepressed K. pneumoniae indicated assembly was rapid for Kp2 and slow for Kpl. Immunoprecipitation of the pulse-chased cell extracts with anti-GroEL antibodies revealed a transient association of newly synthesized NifH and NifDK with GroEL, constituting the first step of nitrogenase assembly. Additional rate-limiting steps are involved in the assembly of Kpl.
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© 1991 Springer Science+Business Media Dordrecht
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Govezensky, D., Greener, T., Zamir, A. (1991). Involvement of GroEL in Klebsiella pneumoniae nif gene expression and nitrogenase assembly. In: Polsinelli, M., Materassi, R., Vincenzini, M. (eds) Nitrogen Fixation. Developments in Plant and Soil Sciences, vol 48. Springer, Dordrecht. https://doi.org/10.1007/978-94-011-3486-6_5
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DOI: https://doi.org/10.1007/978-94-011-3486-6_5
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