title: Mutational analysis of the major soybean UreF paralogue involved in urease activation.
abstract: The soybean genome duplicated ∼14 and 45 million years ago and has many paralogous genes, including those in urease activation (emplacement of Ni and CO(2) in the active site). Activation requires the UreD and UreF proteins, each encoded by two paralogues. UreG, a third essential activation protein, is encoded by the single-copy Eu3, and eu3 mutants lack activity of both urease isozymes. eu2 has the same urease-negative phenotype, consistent with Eu2 being a single-copy gene, possibly encoding a Ni carrier. Unexpectedly, two eu2 alleles co-segregated with missense mutations in the chromosome 2 UreF paralogue (Ch02UreF), suggesting lack of expression/function of Ch14UreF. However, Ch02UreF and Ch14UreF transcripts accumulate at the same level. Further, it had been shown that expression of the Ch14UreF ORF complemented a fungal ureF mutant. A third, nonsense (Q2*) allelic mutant, eu2-c, exhibited 5- to 10-fold more residual urease activity than missense eu2-a or eu2-b, though eu2-c should lack all Ch02UreF protein. It is hypothesized that low-level activation by Ch14UreF is 'spoiled' by the altered missense Ch02UreF proteins ('epistatic dominant-negative'). In agreement with active 'spoiling' by eu2-b-encoded Ch02UreF (G31D), eu2-b/eu2-c heterozygotes had less than half the urease activity of eu2-c/eu2-c siblings. Ch02UreF (G31D) could spoil activation by Chr14UreF because of higher affinity for the activation complex, or because Ch02UreF (G31D) is more abundant than Ch14UreF. Here, the latter is favoured, consistent with a reported in-frame AUG in the 5' leader of Chr14UreF transcript. Translational inhibition could represent a form of 'functional divergence' of duplicated genes.
authors: Biochemistry Department, 117 Schweitzer Hall, University of Missouri, Columbia, MO 65211, USA. polaccoj@missouri.edu
authors: Joe C
authors: JC
authors: Polacco
authors: David L
authors: DL
authors: Hyten
authors: Mônica
authors: M
authors: Medeiros-Silva
authors: David A
authors: DA
authors: Sleper
authors: Kristin D
authors: KD
authors: Bilyeu
mesh_terms: D004252:DNA Mutational Analysis / Q000379:methods*; D009532:Nickel / Q000378:metabolism; D010641:Phenotype; D018515:Plant Leaves / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012639:Seeds / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D013025:Glycine max / Q000201:enzymology* / Q000235:genetics* / Q000378:metabolism; D014510:Urease / Q000235:genetics / Q000378:metabolism*
chemical_list: D010940:Plant Proteins; D009532:Nickel; D014510:Urease