PMID:12384297

Citation	Perlova, O, Nawroth, R, Zellermann, EM and Meletzus, D (2002) Isolation and characterization of the glnD gene of Gluconacetobacter diazotrophicus, encoding a putative uridylyltransferase/uridylyl-removing enzyme. Gene 297:159-68
Abstract	The glnD gene of Gluconacetobacter diazotrophicus was isolated by complementation of the Azotobacter vinelandii glnD (nfrX) mutant strain MV17 using a pLAFR3 cosmid library. The 5 kb chromosomal DNA region encoding the glnD gene on cosmid pAD401 was identified by introduction of deletions as well as subcloning of restriction fragments followed by subsequent DNA sequencing. Three open reading frames were identified with the deduced amino acid sequence of ORF1 showing significant homologies to known GlnD proteins of other proteobacteria such as Sinorhizobium meliloti, Rhizobium tropici, Escherichia coli and Azotobacter vinelandii.A mutagenesis of the chromosomal glnD gene was carried out by insertion of an interposon carrying the kanamycin resistance gene of Tn5. Mutants carrying the cassette inserted into a central region of glnD could not be isolated, while an interposon mutation at the 3' end of glnD was successful. The resulting strain showed a prolonged generation time in complex growth medium and was unable to utilize ammonium as sole nitrogen source. This phenotype appears to be pleiotropic, since the addition of single amino acids to the minimal medium was not sufficient to allow growth. Furthermore, the glnD mutant was able to express nitrogenase under diazotrophic as well as repressing growth conditions.
Links	PubMed
Keywords	Acetobacteraceae/enzymology; Acetobacteraceae/genetics; Amino Acid Sequence; Base Sequence; Blotting, Western; Chromosome Mapping; Cloning, Molecular; DNA, Bacterial/chemistry; DNA, Bacterial/genetics; Gene Expression Regulation, Bacterial; Gene Expression Regulation, Enzymologic; Molecular Sequence Data; Mutagenesis, Insertional; Mutation; Nitrogenase/genetics; Nitrogenase/metabolism; Nucleotidyltransferases/genetics; PII Nitrogen Regulatory Proteins; Sequence Alignment; Sequence Analysis, DNA; Sequence Homology, Amino Acid
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
GLUDI:Q8VS59	involved_in	GO:2001250: positive regulation of ammonia assimilation cycle	ECO:0000315: mutant phenotype evidence used in manual assertion		P		Seeded From UniProt	complete
GLUDI:Q8VS59		GO:2001250: positive regulation of ammonia assimilation cycle	ECO:0000315:		P		Fig 3 - showing glnD mutants were unable to use NH4+ as sole nitrogen source	complete CACAO 8889

References

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