Cloning and characterization of ftsZ and pyrF from the archaeon Thermoplasma acidophilum.
To characterize cytoskeletal components of archaea, the ftsZ gene from Thermoplasma acidophilum was cloned and sequenced. In T. acidophilum ftsZ, which is involved in cell division, was found to be in an operon with the pyrF gene, which encodes orotidine-5'-monophosphate decarboxylase (ODC), an essential enzyme in pyrimidine biosynthesis. Both ftsZ and pyrF from T. acidophilum were expressed in Escherichia coli and formed functional proteins. FtsZ expression in wild-type E. coli resulted in the filamentous phenotype characteristic of ftsZ mutants. T. acidophilum pyrF expression in an E. coli mutant lacking pyrF complemented the mutation and rescued the strain. Sequence alignments of ODCs from archaea, bacteria, and eukarya reveal five conserved regions, two of which have homology to 3-hexulose-6-phosphate synthase (HPS), suggesting a common substrate recognition and binding motif.
Yaoi
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T
NASA Ames Research Center, Moffett Field, California 94035, USA.
Laksanalamai
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P
Jiemjit
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A
Kagawa
H K
HK
Alton
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Trent
J D
JD
D000595:Amino Acid Sequence; D019843:Archaeal Proteins / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D001483:Base Sequence; D001665:Binding Sites; D002455:Cell Division; D003001:Cloning, Molecular; D018387:Codon, Initiator / Q000235:genetics; D017124:Conserved Sequence / Q000235:genetics; D003598:Cytoskeletal Proteins; D004926:Escherichia coli / Q000235:genetics; D005816:Genetic Complementation Test; D008969:Molecular Sequence Data; D009154:Mutation; D009876:Operon / Q000235:genetics; D009964:Orotidine-5'-Phosphate Decarboxylase / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D010641:Phenotype; D011401:Promoter Regions, Genetic / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D015183:Restriction Mapping; D016415:Sequence Alignment; D013822:Thermoplasma / Q000201:enzymology / Q000235:genetics*
D019843:Archaeal Proteins; D001426:Bacterial Proteins; D018387:Codon, Initiator; D003598:Cytoskeletal Proteins; C071525:FtsZ protein, Bacteria; D011994:Recombinant Proteins; C407406:pyrF protein, Archaeal; D009964:Orotidine-5'-Phosphate Decarboxylase