The peptide synthetase catalyzing cyclosporine production in Tolypocladium niveum is encoded by a giant 45.8-kilobase open reading frame.
Cyclosporin A, a potent and clinically-important immunosuppressive drug (SandimmunR), is synthesized from its precursor amino acids by cyclosporin synthetase, a single multi-functional enzyme. In this study we report the cloning of the corresponding coding region of this synthetase. It contains an open reading frame of 45.8 kb which encodes a peptide with a calculated M(r) of 1,689,243. The predicted gene product contains 11 amino-acid-activating domains that are very similar to one another and to the domains of other peptide synthetases. Seven of these domains harbour N-methyltransferase functions. This is the largest genomic ORF described so far.
Weber
G
G
Biochemie Ges.m.b.H. Kufstein-Schaftenau, Austria.
Schörgendorfer
K
K
Schneider-Scherzer
E
E
Leitner
E
E
D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D003001:Cloning, Molecular; D016384:Consensus Sequence; D016572:Cyclosporine / Q000096:biosynthesis*; D003904:Mitosporic Fungi / Q000201:enzymology / Q000235:genetics*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009097:Multienzyme Complexes / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D009838:Oligodeoxyribonucleotides; D016366:Open Reading Frames; D010453:Peptide Synthases / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D015183:Restriction Mapping; D017386:Sequence Homology, Amino Acid
D009097:Multienzyme Complexes; D009838:Oligodeoxyribonucleotides; C048525:cyclosporin A synthetase; D016572:Cyclosporine; D010453:Peptide Synthases