A trans-acting gene is required for the phenotypic expression of a tyrosinase gene in Streptomyces.
The melanin locus (melC) from Streptomyces antibioticus was previously shown to be composed of two open reading frames (ORFs), melC1 and melC2. The melC2 ORF codes for the polypeptide chain of tyrosinase (apotyrosinase). The function of melC1 is not known except that insertional mutation within it abolishes the tyrosinase activity. Here, we show that in Streptomyces lividans TK64 harboring melC1 mutated and melC2 intact (melC1- melC2+) plasmids, while there was no tyrosinase activity, melC transcript was synthesized and apotyrosinase could be detected. The apotyrosinase could be activated to a limited degree by incubation with copper ions, or by mixing the mycelial extract from a culture harboring a melC1- melC2+ (pPF950) plasmid with that from a culture containing a melC1+ melC2- (pSA1) plasmid. Complementation analysis showed that melC1 acted in trans on the tyrosinase gene expression. Together, these results suggest that melC1 encodes or regulates a copper-transfer protein serving an in vivo copper-donor function in the biosynthesis of active tyrosinase.
Lee
Y H
YH
Institute of Biochemistry, National Yang-Ming Medical College, Taipei, Taiwan.
Chen
B F
BF
Wu
S Y
SY
Leu
W M
WM
Lin
J J
JJ
Chen
C W
CW
Lo
S C
SC
D004156:Catechol Oxidase / Q000235:genetics*; D004262:DNA Restriction Enzymes; D005796:Genes; D005798:Genes, Bacterial; D014442:Monophenol Monooxygenase / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D009154:Mutation; D009693:Nucleic Acid Hybridization; D010641:Phenotype; D010957:Plasmids; D013302:Streptomyces / Q000201:enzymology / Q000235:genetics*; D014158:Transcription, Genetic
D004156:Catechol Oxidase; D014442:Monophenol Monooxygenase; D004262:DNA Restriction Enzymes