title: Activation of the urease of Schizosaccharomyces pombe by the UreF accessory protein from soybean.
abstract: Plant orthologs of the bacterial urease accessory genes ureD and ureF, which are required for the insertion of the nickel ion at the active site, have been isolated from soybean ( Glycine max L. Merr.), tomato ( Lycopersicon esculentum) and Arabidopsis thaliana. The functionality of soybean UreD and UreF was tested by measuring their ability to complement urease-negative mutants of Schizosaccharomyces pombe, a eukaryote which produces a "plant-like" urease of ~90 kDa. The S. pombe ure4 mutant was complemented by a 12-kb fragment of S. pombe genomic DNA, which was shown by PCR to contain a putative ureD gene. However, ure4 was not complemented by a UreD cDNA soybean, expressed under the control of a strong promoter. In contrast, an S. pombe ure3 mutation was complemented by both a 10-kb fragment of S. pombe DNA containing ureF and the UreF cDNA from soybean. Soybean Eu2 is a candidate urease accessory gene; its product cooperates with the Eu3 protein in activating apourease in vitro. However, the sequences of UreD and UreF transcripts from two eu2/eu2 mutants, recovered as RT-PCR products, revealed no mutational alteration, suggesting that Eu2 encodes neither UreD nor UreF.
authors.affiliation: Department of Biochemistry and Interdisciplinary Plant Group, 117 Schweitzer Hall, University of Missouri-Columbia, Columbia, MO 65211, USA.
authors.forename: M
authors.initials: M
authors.lastname: Bacanamwo
authors.forename: C-P
authors.initials: CP
authors.lastname: Witte
authors.forename: M W
authors.initials: MW
authors.lastname: Lubbers
authors.forename: J C
authors.initials: JC
authors.lastname: Polacco
mesh_terms: D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000201:enzymology / Q000235:genetics; D001426:Bacterial Proteins / Q000502:physiology; D002352:Carrier Proteins / Q000502:physiology; D017931:DNA Primers / Q000737:chemistry; D004789:Enzyme Activation; D015964:Gene Expression Regulation, Bacterial; D005816:Genetic Complementation Test; D066298:In Vitro Techniques; D018551:Solanum lycopersicum / Q000235:genetics; D008969:Molecular Sequence Data; D009532:Nickel; D028044:Phosphate-Binding Proteins; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012568:Schizosaccharomyces / Q000201:enzymology*; D017386:Sequence Homology, Amino Acid; D013025:Glycine max / Q000201:enzymology*; D014510:Urease / Q000235:genetics / Q000378:metabolism* / Q000502:physiology*
chemical_list: D001426:Bacterial Proteins; D002352:Carrier Proteins; D017931:DNA Primers; D028044:Phosphate-Binding Proteins; C086767:UreD protein, Bacteria; C101639:UreF protein, bacteria; C082270:ureG protein, Bacteria; D009532:Nickel; D014510:Urease