title: The soybean Eu3 gene encodes an Ni-binding protein necessary for urease activity.
abstract: Mutation in Eu3 eliminates activity of both soybean ureases, the embryo-specific (encoded by Eu1) and the tissue-ubiquitous (encoded by Eu4). eu3-e1 is a completely recessive null allele. Eu3-e3 is a semi-dominant specifying 0.1% wild-type urease activity in the homozygous state and 5-10% as a heterozygote (Meyer-Bothling et al. 1987). Antibodies to plant UreG, a homologue of the bacterial urease accessory protein, revealed a 32 kDa protein (p32) in embryos of the Eu3/Eu3 precursor genotype. p32 is identical to UreG by the criteria of size, antigenicity, and its ability to bind Ni2+, a trait expected from the deduced histidine-rich N-terminus of UreG. UreG was absent in eu3-e1/eu3-e1, and lack of UreG co-segregated with eu3-e1. Eu3-e3 specified a UreG transcript which coded valine in place of alanine at residue 142 (A142V) confirming thatEu3 encodes UreG, which is renamed Eu3. Eu3 (A142V) retained Ni-binding ability. Eu3 is directly involved in urease activation, since anti-Eu3 (UreG) antibodies inhibited the in vitro activation of urease. Eu1 (embryo urease) and Eu3 accumulated in parallel in the developing embryo. The presence of Eu1 was not necessary for the high embryonic level of Eu3. However, the presence of Eu3 appeared to be important for accumulation of Eu1, perhaps by stabilizing it by Ni insertion. At the level of sensitivity employed Eu3 was detected in crude extracts of embryos but not non-embryonic tissues which have 1/500th the embryo urease activity. Functional Eu3, however, is necessary for activation of the ubiquitous urease in non-embryonic tissues.
authors.affiliation: Department of Biochemistry and Interdisciplinary Plant Group, 117 Schweitzer Hall, University of Missouri, Columbia, MO 65211, USA.
authors.forename: S K
authors.initials: SK
authors.lastname: Freyermuth
authors.forename: M
authors.initials: M
authors.lastname: Bacanamwo
authors.forename: J C
authors.initials: JC
authors.lastname: Polacco
mesh_terms: D000595:Amino Acid Sequence; D002352:Carrier Proteins / Q000235:genetics*; D018076:DNA, Complementary; D004789:Enzyme Activation; D020558:GTP Phosphohydrolases; D018507:Gene Expression Regulation, Developmental; D018506:Gene Expression Regulation, Plant; D008969:Molecular Sequence Data; D009532:Nickel / Q000378:metabolism*; D009711:Nucleotides / Q000378:metabolism; D011485:Protein Binding; D017386:Sequence Homology, Amino Acid; D030262:Soybean Proteins; D013025:Glycine max / Q000235:genetics*; D014510:Urease / Q000378:metabolism*
chemical_list: D002352:Carrier Proteins; D018076:DNA, Complementary; D009711:Nucleotides; D030262:Soybean Proteins; D009532:Nickel; D014510:Urease; D020558:GTP Phosphohydrolases; C406502:ureG protein, Glycine max