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PMID: 8121412 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

A 4.6 kb DNA region of Rhizobium meliloti involved in determining urease and hydrogenase activities carries the structural genes for urease (ureA, ureB, ureC) interrupted by other open reading frames.

Molecular & general genetics : MGG ·Vol. 242 ·No. 5 ·1994-03-00 ·Pages 539-50

Miksch G, Arnold W, Lentzsch P, Priefer UB, Pühler A

Abstract

A 4.6 kb DNA region of the Rhizobium meliloti strain AK631 was found to contain seven open reading frames (ORFs), all oriented in the same direction. The putative gene products of four of these ORFs were highly homologous to UreA, UreB and UreC of Klebsiella aerogenes, Proteus mirabilis, Proteus vulgaris and Canavalia ensiformis. The overall organisation of the DNA region analysed was ORF1, ureA (ORF2), ORF3, ureB (ORF4), ORF5, ORF6 and ureC (ORF7), indicating that the organisation of the urease structural genes in R. meliloti differs from that of other urease genes so far characterized. ORF1 was incomplete; only the 3' end of the coding region was present. The six complete ORFs coded for polypeptides of 11.1 (UreA), 8.9 (ORF3), 10.8 (UreB), 15.0 (ORF5), 13.8 (ORF6) and 60.7 kDa (UreC). No sequence homology to known polypeptides could be detected for the gene products of ORF1, ORF3, ORF5 and ORF6. Using a lacZ fusion and insertional mutagenesis it was shown that the seven ORFs identified were all located in the same transcription unit. For mutational analysis a resistance gene cassette was introduced into each of the complete ORFs resulting in apolar mutations. Mutations in ureA, ureB and ureC, but not in ORF3, ORF5 and ORF6, abolished urease activity in R. meliloti. The determination of hydrogen uptake in these R. meliloti mutants revealed that only ORF6 and ureB are necessary for hydrogen uptake.

Related Genes
f1p6A ureB ureC
MeSH Terms
Amino Acid Sequence Base Sequence Binding Sites Cloning, Molecular DNA Mutational Analysis Genes, Plant Hydrogenase/genetics Molecular Sequence Data Mutagenesis, Insertional Open Reading Frames Sinorhizobium meliloti/genetics Urease/genetics
Chemicals
Hydrogenase Urease
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Miksch G
Lehrstuhl für Genetik, Fakultät für Biologie, Universität Bielefeld, Germany.
Arnold W
Lentzsch P
Priefer U B
Pühler A
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Article Info
Journal
Molecular & general genetics : MGG
Abbr.
Mol Gen Genet
ISSN
0026-8925
DOI
10.1007/BF00285277
Published
1994-03-00
Pages
539-50
Language
English
Region
Germany
NLM ID
0125036
Subset
IM
Databases
GENBANK
S69145
External Links
PubMed source DOI: 10.1007/BF00285277
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