Temperature-sensitive, osmotic-remedial mutants of Neurospora crassa: osmotic pressure induced alterations of enzyme stability

Abstract

Three temperature-sensitive, adenine-requiring mutants of Neurospora crassa were found to be osmotic-remedial when non-penetrating solutes were used to increase the osmolarity of the growth medium. The affected enzyme (adenylosuccinase) from one of the mutants (ad-4, 44206t) was found to have higher levels of activity when the organism was grown at non-permissive temperatures under osmotic-remedial conditions than when it was grown with adenine as a nutritional supplement. The enzyme synthesized at 30 degrees C in the presence of adenine exhibited increased sensitivity to inhibition by high salt concentrations and a lowered stability toward heat denaturation, indicating that the remedial effect may be the result of changes in the physical properties of the enzyme molecule. Temperature shift experiments indicate that the enzyme which is synthesized at permissive temperatures or under osmotic-remedial conditions is also stable in vivo under non-permissive conditions. This suggests that the critical period for temperature sensitivity, and conversely osmotic remediability, may be during protein synthesis or during the conformational folding of the protein.