Activation of the urease of Schizosaccharomyces pombe by the UreF accessory protein from soybean.
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.
Bacanamwo
M
M
Department of Biochemistry and Interdisciplinary Plant Group, 117 Schweitzer Hall, University of Missouri-Columbia, Columbia, MO 65211, USA.
Witte
C-P
CP
Lubbers
M W
MW
Polacco
J C
JC
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*
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