title: Modified pathway to synthesize ribulose 1,5-bisphosphate in methanogenic archaea.
abstract: Several sequencing projects unexpectedly uncovered the presence of genes that encode ribulose 1,5-bisphosphate (RuBP) carboxylase/oxygenase (RubisCO) in anaerobic archaea. RubisCO is the key enzyme of the Calvin-Benson-Bassham (CBB) reductive pentose phosphate pathway, a scheme that does not appear to contribute greatly, if at all, to net CO2 assimilation in these organisms. Recombinant forms of the archaeal enzymes do, however, catalyze a bona fide RuBP-dependent CO2 fixation reaction, and it was recently shown that Methanocaldococcus (Methanococcus) jannaschii and other anaerobic archaea synthesize catalytically active RubisCO in vivo. To complete the CBB pathway, there is a need for an enzyme, i.e., phosphoribulokinase (PRK), to catalyze the formation of RuBP, the substrate for the RubisCO reaction. Homology searches, as well as direct enzymatic assays with M. jannaschii, failed to reveal the presence of PRK. The apparent lack of PRK raised the possibility that either there is an alternative pathway to generate RuBP or RubisCO might use an alternative substrate in vivo. In the present study, direct enzymatic assays performed with alternative substrates and extracts of M. jannsachii provided evidence for a previously uncharacterized pathway for RuBP synthesis from 5-phospho-D-ribose-1-pyrophosphate (PRPP) in M. jannaschii and other methanogenic archaea. Proteins and genes involved in the catalytic conversion of PRPP to RuBP were identified in M. jannaschii (Mj0601) and Methanosarcina acetivorans (Ma2851), and recombinant Ma2851 was active in extracts of Escherichia coli. Thus, in this work we identified a novel means to synthesize the CO2 acceptor and substrate for RubisCO in the absence of a detectable kinase, such as PRK. We suggest that the conversion of PRPP to RuBP might be an evolutional link between purine recycling pathways and the CBB scheme.
authors.affiliation: Department of Microbiology, The Ohio State University, Columbus, Ohio 43210-1292, USA.
authors.forename: Michael W
authors.initials: MW
authors.lastname: Finn
authors.forename: F Robert
authors.initials: FR
authors.lastname: Tabita
mesh_terms: D002245:Carbon Dioxide / Q000378:metabolism; D002384:Catalysis; D017017:Methanococcus / Q000378:metabolism*; D009243:NAD / Q000494:pharmacology; D010427:Pentose Phosphate Pathway; D011755:Pyrophosphatases / Q000378:metabolism; D012274:Ribulosephosphates / Q000096:biosynthesis*
chemical_list: D012274:Ribulosephosphates; D009243:NAD; D002245:Carbon Dioxide; C001933:ribulose-1,5 diphosphate; D011755:Pyrophosphatases