Cloning and Characterization of ftsZ and pyrF from the Archaeon Thermoplasma acidophilum

• Published in: Biochemical and biophysical research communications Vol. 275; no. 3; pp. 936 - 945
• Main Authors: Yaoi, Takuro, Laksanalamai, Pongpan, Jiemjit, Anchalee, Kagawa, Hiromi K., Alton, Thomas, Trent, Jonathan D.
• Format: Journal Article
• Language: English
• Published: Legacy CDMS Elsevier Inc 07-09-2000
• Subjects: Amino Acid Sequence; archaea; Archaeal Proteins - chemistry; Archaeal Proteins - genetics; Archaeal Proteins - metabolism; Bacterial Proteins - chemistry; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Base Sequence; Binding Sites; Cell Division; Cloning, Molecular; Codon, Initiator - genetics; Conserved Sequence - genetics; Cytoskeletal Proteins; cytoskeleton; Escherichia coli; Escherichia coli - genetics; ftsZ gene; Genetic Complementation Test; genetics; Life Sciences (General); molecular evolution; Molecular Sequence Data; Mutation; Operon - genetics; orotidine 5'-monophosphate decarboxylase; Orotidine-5'-Phosphate Decarboxylase - chemistry; Orotidine-5'-Phosphate Decarboxylase - genetics; Orotidine-5'-Phosphate Decarboxylase - metabolism; Phenotype; Promoter Regions, Genetic - genetics; pyrF gene; Recombinant Proteins - chemistry; Recombinant Proteins - metabolism; Restriction Mapping; Sequence Alignment; Space life sciences; Thermoplasma - enzymology; Thermoplasma - genetics; Thermoplasma acidophilum; cell division; genetics; cytoskeleton; molecular evolution; archaea
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1006/bbrc.2000.3401

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.