The adenylosuccinate synthetase from the hyperthermophilic archaeon Pyrococcus species displays unusual structural features.
The first example of a hyperthermophilic adenylosuccinate synthetase is reported, which is an enzyme that must maintain its folded structure at temperatures as high as 102 degrees C. The amino acid sequence of this key enzyme has been determined after cloning and sequencing the purA-like gene from the archaeal Pyrococcus sp. strain ST700. The corresponding protein displays two unexpected features: (1) it is 21% shorter than the homologous mesophilic enzymes and this shortening corresponds to the loss of two alpha-helices and three beta-strands present in the Escherichia coli enzyme; (2) surprisingly, the archaeal adenylosuccinate synthetase has a significant number of substitutions in residues that are conserved in all other homologous enzymes from bacteria to man. In E. coli, the conserved residues have been described as essential for catalytic activity and/or for maintaining the folded structure of the homodimer. Despite these drastic differences, the purA-like archaeal gene seems to be normally expressed and its product functions in vivo in bacteria, since it complemented an E. coli purA auxotroph. The archaeal adenylosuccinate synthetase appears to be a good example of a bona fide orthologous protein. Reconstruction of phylogenetic trees showed that the archaeal gene is equally distantly related to both eukaryotes and bacteria, independently of the numerous substitutions observed at critical positions.
Bouyoub
A
A
Institut de Génétique et Microbiologie, URA 1354, Université Paris-Sud, Orsay, France.
Barbier
G
G
Forterre
P
P
Labedan
B
B
D000265:Adenylosuccinate Synthase / Q000737:chemistry / Q000235:genetics*; D000595:Amino Acid Sequence; D001105:Archaea / Q000201:enzymology* / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D003850:Deoxyribonuclease I / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial / Q000235:genetics; D005816:Genetic Complementation Test; D008969:Molecular Sequence Data; D010802:Phylogeny; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
D003850:Deoxyribonuclease I; D000265:Adenylosuccinate Synthase