ATP-induced protyrosinase synthesis and carboxyl-terminal processing in Neurospora crassa.
The effects of 3'-5' cyclic AMP and ATP upon tyrosinase induction in Neurospora crassa were examined. Northern analysis of total cellular RNA revealed rapid de novo synthesis of protyrosinase after addition of these substances to stationary-phase mycelia. The maturation of protyrosinase in crude extracts of mycelia was followed by Western analysis. Polyclonal rabbit antiserum directed against the denatured carboxyl-terminal extension of protyrosinase does recognize the proform and several intermediate forms of different molecular weight but not mature tyrosinase. Disruption of ATP-induced mycelia in sodium phosphate buffer (pH 6.0) demonstrate processing at the carboxyl-terminal end of protyrosinase. The activity assays revealed that protyrosinase is an inactive precursor and that at least two active forms of slightly different molecular weight are present in crude extracts. Maturation of protyrosinase thus involves specific and sequential proteolytic cleavage at the carboxyl-terminus. These results suggest the presence of a tyrosinase activator in Neurospora crassa mycelia, which is kept apart from protyrosinase in the intact mycelium.
Kupper
U
U
Biochemisches Institut, Universität Zürich, Switzerland.
Niedermann
D M
DM
Schilling
B C
BC
Lerch
K
K
D000255:Adenosine Triphosphate / Q000502:physiology*; D015153:Blotting, Western; D004789:Enzyme Activation; D004792:Enzyme Precursors / Q000096:biosynthesis*; D007700:Kinetics; D014442:Monophenol Monooxygenase / Q000378:metabolism*; D009492:Neurospora crassa / Q000201:enzymology*; D011499:Protein Processing, Post-Translational; D012331:RNA, Fungal / Q000096:biosynthesis; D012333:RNA, Messenger / Q000096:biosynthesis
D004792:Enzyme Precursors; D012331:RNA, Fungal; D012333:RNA, Messenger; D000255:Adenosine Triphosphate; D014442:Monophenol Monooxygenase