Cloning and expression of the S-adenosylmethionine decarboxylase gene of Neurospora crassa and processing of its product.
S-adenosylmethionine decarboxylase (AdoMetDC) catalyzes the formation of decarboxylated AdoMetDC, a precursor of the polyamines spermidine and spermine. The enzyme is derived from a proenzyme by autocatalytic cleavage. We report the cloning and regulation of the gene for AdoMetDC in Neurospora crassa, spe-2, and the effect of putrescine on enzyme maturation and activity. The gene was cloned from a genomic library by complementation of a spe-2 mutant. Like other AdoMetDCs, that of Neurospora is derived by cleavage of a proenzyme. The deduced sequence of the Neurospora proenzyme (503 codons) is over 100 codons longer than any other AdoMetDC sequence available in genomic databases. The additional amino acids are found only in the AdoMetDC of another fungus, Aspergillus nidulans, a cDNA for which we also sequenced. Despite the conserved processing site and four acidic residues required for putrescine stimulation of human proenzyme processing, putrescine has no effect on the rate (t0.5 approximately 10 min) of processing of the Neurospora gene product. However, putrescine is absolutely required for activity of the Neurospora enzyme (K0.5 approximately 100 microM). The abundance of spe-2 mRNA and enzyme activity is regulated 2- to 4-fold by spermidine.
Hoyt
M A
MA
Department of Molecular Biology and Biochemistry, University of California at Irvine 92697-3900, USA.
Williams-Abbott
L J
LJ
Pitkin
J W
JW
Davis
R H
RH
D012437:Adenosylmethionine Decarboxylase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D000595:Amino Acid Sequence; D001233:Aspergillus nidulans / Q000235:genetics; D003001:Cloning, Molecular; D003062:Codon / Q000235:genetics; D017124:Conserved Sequence; D016208:Databases, Factual; D005800:Genes, Fungal; D005816:Genetic Complementation Test; D015698:Genomic Library; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D009492:Neurospora crassa / Q000201:enzymology* / Q000235:genetics*; D012150:Polymorphism, Restriction Fragment Length; D011700:Putrescine / Q000494:pharmacology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism; D015183:Restriction Mapping; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D003062:Codon; D011994:Recombinant Proteins; D012437:Adenosylmethionine Decarboxylase; D011700:Putrescine