Coproporphyrin excretion and low thiol levels caused by point mutation in the Rhodobacter sphaeroides S-adenosylmethionine synthetase gene.
A spontaneous mutant of Rhodobacter sphaeroides f. sp. denitrificans IL-106 was found to excrete a large amount of a red compound identified as coproporphyrin III, an intermediate in bacteriochlorophyll and heme synthesis. The mutant, named PORF, is able to grow under phototrophic conditions but has low levels of intracellular cysteine and glutathione and overexpresses the cysteine synthase CysK. The expression of molybdoenzymes such as dimethyl sulfoxide (DMSO) and nitrate reductases is also affected under certain growth conditions. Excretion of coproporphyrin and overexpression of CysK are not directly related but were both found to be consequences of a diminished synthesis of the key metabolite S-adenosylmethionine (SAM). The wild-type phenotype is restored when the gene metK encoding SAM synthetase is supplied in trans. The metK gene in the mutant strain has a mutation leading to a single amino acid change (H145Y) in the encoded protein. This point mutation is responsible for a 70% decrease in intracellular SAM content which probably affects the activities of numerous SAM-dependent enzymes such as coproporphyrinogen oxidase (HemN); uroporphyrinogen III methyltransferase (CobA), which is involved in siroheme synthesis; and molybdenum cofactor biosynthesis protein A (MoaA). We propose a model showing that the attenuation of the activities of SAM-dependent enzymes in the mutant could be responsible for the coproporphyrin excretion, the low cysteine and glutathione contents, and the decrease in DMSO and nitrate reductase activities.
Sabaty
Monique
M
Laboratoire de Bioénergétique Cellulaire, CEA CADARACHE, DSV, IBEB, Saint-Paul-lez-Durance, F-13108, France. monique.sabaty@cea.fr
Adryanczyk
Géraldine
G
Roustan
Chloë
C
Cuiné
Stephan
S
Lamouroux
Christine
C
Pignol
David
D
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution / Q000235:genetics; D003306:Coproporphyrins / Q000378:metabolism*; D003545:Cysteine / Q000378:metabolism; D004252:DNA Mutational Analysis; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D005816:Genetic Complementation Test; D005978:Glutathione / Q000378:metabolism; D008716:Methionine Adenosyltransferase / Q000235:genetics* / Q000378:metabolism*; D008954:Models, Biological; D008969:Molecular Sequence Data; D020125:Mutation, Missense; D017354:Point Mutation; D012242:Rhodobacter sphaeroides / Q000201:enzymology* / Q000235:genetics* / Q000378:metabolism; D012436:S-Adenosylmethionine / Q000378:metabolism; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D013438:Sulfhydryl Compounds / Q000378:metabolism*
D003306:Coproporphyrins; D004269:DNA, Bacterial; D013438:Sulfhydryl Compounds; D012436:S-Adenosylmethionine; D008716:Methionine Adenosyltransferase; D005978:Glutathione; D003545:Cysteine