Functional knockout of the adenosine 5'-phosphosulfate reductase gene in Physcomitrella patens revives an old route of sulfate assimilation.
The reduction of adenosine 5'-phosphosulfate (APS) to sulfite catalyzed by adenosine 5'-phosphosulfate reductase is considered to be the key step of sulfate assimilation in higher plants. However, analogous to enteric bacteria, an alternative pathway of sulfate reduction via phosphoadenosine 5'-phosphosulfate (PAPS) was proposed. To date, the presence of the corresponding enzyme, PAPS reductase, could be neither confirmed nor excluded in plants. To find possible alternative routes of sulfate assimilation we disrupted the adenosine 5'-phosphosulfate reductase single copy gene in Physcomitrella patens by homologous recombination. This resulted in complete loss of the correct transcript and enzymatic activity. Surprisingly, the knockout plants grew on sulfate as the sole sulfur source, and the concentration of thiols in the knockouts did not differ from the wild type plants. However, when exposed to a sublethal concentration of cadmium, the knockouts were more sensitive than wild type plants. When fed [(35)S]sulfate, the knockouts incorporated (35)S in thiols; the flux through sulfate reduction was approximately 50% lower than in the wild type plants. PAPS reductase activity could not be measured with thioredoxin as reductant, but a cDNA and a gene coding for this enzyme were detected in P. patens. The moss Physcomitrella patens is thus the first plant species wherein PAPS reductase was confirmed on the molecular level and also the first organism wherein both APS- and PAPS-dependent sulfate assimilation co-exist.
Koprivova
Anna
A
Department of Plant Biotechnology, University of Freiburg, D-79104 Freiburg, Germany.
Meyer
Andreas J
AJ
Schween
Gabriele
G
Herschbach
Cornelia
C
Reski
Ralf
R
Kopriva
Stanislav
S
D000250:Adenosine Phosphosulfate / Q000378:metabolism*; D000595:Amino Acid Sequence; D001483:Base Sequence; D019068:Bryopsida / Q000201:enzymology* / Q000235:genetics; D003001:Cloning, Molecular; D017931:DNA Primers; D017353:Gene Deletion; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D050862:Oxidoreductases Acting on Sulfur Group Donors; D030821:Plants, Genetically Modified; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013431:Sulfates / Q000378:metabolism*
D017931:DNA Primers; D013431:Sulfates; D000250:Adenosine Phosphosulfate; D010088:Oxidoreductases; D050862:Oxidoreductases Acting on Sulfur Group Donors; C019618:adenylylsulfate reductase