Functional characterisation of urease accessory protein G (ureG) from potato.
The activation of the nickel metalloenzyme urease is a complex process. In bacteria, several urease accessory proteins are essential for incorporation of nickel into the active centre of urease. Comparatively little is known about the activation process and the proteins involved in plants. We cloned five different cDNAs encoding isoforms of urease accessory protein G (ureG) in potato. The 5'-coding region of these cDNAs is highly polymorphic within Solanum tuberosum ssp. tuberosum, containing mainly a simple sequence repeat encoding histidine and aspartate. Mapping on an ultrahigh-density map of the potato genome and Southern blot analysis showed that the isoforms arise from allelic differences of a single-copy gene which was located on chromosome 2. Expression analysis at the mRNA and protein levels indicated the presence of ureG in almost all tissues examined, consistent with the ubiquitous expression of urease. An attempt to correlate urease activity with ureG expression levels in different tissues was made. Allelic copies of ureG were expressed in a tissue-specific manner. UreG from potato and the Klebsiella aerogenes urease operon defective in bacterial ureG were co-expressed in Escherichia coli. The plant gene complements the K. aerogenes ureG mutation, demonstrating that it encodes a urease accessory protein and indicating a structural conservation between the plant and the bacterial urease activation complexes.
Witte
C P
CP
Unit of Plant Biochemistry, Scottish Crop Research Institute, Invergowrie, Dundee , UK. cwitte@scri.sari.ac.uk
Isidore
E
E
Tiller
S A
SA
Davies
H V
HV
Taylor
M A
MA
D000483:Alleles; D000595:Amino Acid Sequence; D001483:Base Sequence; D015139:Blotting, Southern; D002352:Carrier Proteins / Q000235:genetics*; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D018744:DNA, Plant / Q000235:genetics; D021902:Enterobacter aerogenes / Q000235:genetics; D020558:GTP Phosphohydrolases; D018506:Gene Expression Regulation, Plant; D005816:Genetic Complementation Test; D008969:Molecular Sequence Data; D009154:Mutation; D009876:Operon; D020033:Protein Isoforms / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid; D011198:Solanum tuberosum / Q000201:enzymology / Q000235:genetics*; D030262:Soybean Proteins; D014018:Tissue Distribution; D014510:Urease / Q000235:genetics / Q000378:metabolism
D002352:Carrier Proteins; D018076:DNA, Complementary; D018744:DNA, Plant; D020033:Protein Isoforms; D012333:RNA, Messenger; D030262:Soybean Proteins; D014510:Urease; D020558:GTP Phosphohydrolases; C406502:ureG protein, Glycine max