Effects of temperature shifts on the activities of Neurospora crassa glycogen synthase, glycogen phosphorylase and trehalose-6-phosphate synthase.
Conidiospore germlings of Neurospora crassa submitted to a heat shock at 45 degrees C accumulate trehalose and degrade glycogen. The opposite occurs upon reincubation at a physiologic temperature (30 degrees C). These observations suggest a temperature-dependent mechanism for the preferential synthesis of one or the other sugar reserve. Here we show that concomitant with these shifts of temperature, occurred reversible changes in the activities of glycogen synthase and phosphorylase. Glycogen synthase was inactivated at 45 degrees C while phosphorylase was activated. The reverse was true when the cells were shifted back to 30 degrees C. Addition of cycloheximide did not prevent the reversible enzymatic changes, which remained stable after gel filtration. Apparently, the effects of temperature shifts occurred at the level of reversible covalent enzymatic modifications. Trehalose-6-phosphate synthase properties were also affected by temperature. For instance, the enzyme was less sensitive to in vitro inhibition by inorganic phosphate at 50 degrees C than at 30 degrees C. Fructose-6-phosphate partially relieved the inhibitory effect of phosphate at 30 degrees C but not at 50 degrees C. These effects of the assay temperature, inorganic phosphate, and fructose-6-phosphate, on trehalose-6-phosphate synthase activity, were more evident for crude extracts obtained from heat-shocked cells. Altogether, these results may contribute to explain the preferential accumulation of trehalose 45 degrees C, or that of glycogen at 30 degrees C.
Noventa-Jordão
M A
MA
Departamento de Biologia, Faculdade de Filosofia, Universidade de São Paulo, Ribeirão Preto, Brasil.
de Lourdes
M
M
Polizeli
T M
TM
Bonini
B M
BM
Jorge
J A
JA
Terenzi
H F
HF
D003513:Cycloheximide / Q000494:pharmacology; D005636:Fructosephosphates / Q000494:pharmacology; D005964:Glucosyltransferases / Q000378:metabolism*; D006003:Glycogen / Q000378:metabolism; D006006:Glycogen Synthase / Q000378:metabolism*; D006358:Hot Temperature; D009492:Neurospora crassa / Q000201:enzymology*; D010710:Phosphates / Q000494:pharmacology; D006005:Phosphorylases / Q000378:metabolism*; D014199:Trehalose / Q000378:metabolism
D005636:Fructosephosphates; D010710:Phosphates; C027618:fructose-6-phosphate; D006003:Glycogen; D003513:Cycloheximide; D014199:Trehalose; D005964:Glucosyltransferases; D006005:Phosphorylases; D006006:Glycogen Synthase; C023180:trehalose-6-phosphate synthase