Human molybdopterin synthase gene: identification of a bicistronic transcript with overlapping reading frames.
A universal molybdenum-containing cofactor (MoCo) is essential for the activity of all human molybdoenzymes, including sulphite oxidase. The free cofactor is highly unstable, and all organisms share a similar biosynthetic pathway. The involved enzymes exhibit homologies, even between bacteria and humans. We have exploited these homologies to isolate a cDNA for the heterodimeric molybdopterin (MPT)-synthase. This enzyme is necessary for the conversion of an unstable precursor into molybdopterin, the organic moiety of MoCo. The corresponding transcript shows a bicistronic structure, encoding the small and large subunits of the MPT-synthase in two different open reading frames (ORFs) that overlap by 77 nucleotides. In various human tissues, only one size of mRNA coinciding with the bicistronic transcript was detected. In vitro translation and mutagenesis experiments demonstrated that each ORF is translated independently, leading to the synthesis of a 10-kDa protein and a 21-kDa protein for the small and large subunits, respectively, and indicated that the 3'-proximal ORF of the bicistronic transcript is translated by leaky scanning.
Botanisches Institut der Technischen Universität, Braunschweig, Humboldtstr. 1, D-38106 Braunschweig, Germany.
B
B
Stallmeyer
G
G
Drugeon
J
J
Reiss
A L
AL
Haenni
R R
RR
Mendel
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D003067:Coenzymes; D004926:Escherichia coli / Q000235:genetics; D015723:Gene Library; D005807:Genes, Overlapping; D006160:Guanosine Triphosphate / Q000378:metabolism; D006801:Humans; D008099:Liver / Q000378:metabolism; D008667:Metalloproteins / Q000378:metabolism; D008969:Molecular Sequence Data; D000090006:Molybdenum Cofactors; D016254:Mutagenesis, Insertional; D014176:Protein Biosynthesis / Q000502:physiology; D011621:Pteridines / Q000378:metabolism; D011817:Rabbits; D016364:Reading Frames / Q000235:genetics; D012156:Reticulocytes / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013466:Sulfurtransferases / Q000235:genetics*
D003067:Coenzymes; D006160:Guanosine Triphosphate; D008667:Metalloproteins; D000090006:Molybdenum Cofactors; D011621:Pteridines; D013466:Sulfurtransferases; C022261:molybdopterin; C085585:molybdopterin synthase