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Microbiology 144 (1998), 1537-1547; DOI  10.1099/00221287-144-6-1537
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Characterization of the glnB gene product of Nostoc punctiforme strain ATCC 29133: glnB or the PII protein may be essential

Thomas E. Hanson1, Karl Forchhammer2, Nicole Tandeau de Marsac3 and John C. Meeks1,*

1 Section of Microbiology, University of California, One Shields Avenue, Davis, CA 95616, USA
2 Lehrstuhl für Mikrobiologie der Universität München, Maria-Ward-Str. 1a, D-80638 München, Germany
3 Unité de Physiologie Microbienne, Département de Biochimie et Génétique Moléculaire, Institut Pasteur, 28 rue du Docteur Roux, 75724 Paris Cedex 15, France

ABSTRACT

Summary: Bacterial PII proteins, encoded by glnB genes, are central signalling molecules in nitrogen regulatory pathways and are modulated by post-translational modification in response to the cellular nitrogen status. The glnB gene was cloned from the filamentous heterocyst-forming cyanobacterium Nostoc punctiforme strain ATCC 29133 (PCC 73102) by heterologous hybridization to a Synechococcus sp. strain PCC 7942 gene fragment. Expression of the cloned gene was verified by hybridization to N. punctiforme total RNA and a single cross-reactive polypeptide was observed in immunoblots of N. punctiforme extracts probed with anti-Synechococcus 7942 PII antiserum. Modification of the purified N. punctiforme PII protein by a Synechococcus 7942 PII kinase was observed, but modified forms of PII were not detected in extracts of N. punctiforme from a variety of incubation conditions. The N. punctiforme glnB gene could not be disrupted by targeted gene replacement unless a second copy of glnB was provided in trans, suggesting that the gene or gene product is essential for growth under the conditions tested.

* Author for correspondence: John C. Meeks. Tel: + 1 530 752 3346. Fax: +1 530 752 9014. e-mail: jcmeeks@ucdavis.edu

Keywords: Nostoc punctiforme,, diazotrophic cyanobacterium, glnB gene, PII protein




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