Nocodazole-Resistant Mutants in Paramecium

The effect of the microtubule inhibitor nocodazole was studied on Paramecium and shown to arrest cell multiplication, depolymerize the internal microtubule network, and block the development of macro- and micronuclear spindles and of the cytospindle (a cortical microtubule array assembled during division). After ultraviolet mutagenesis, three mutants resistant to nocodazole, that is capable of continued growth in the presence of the drug, were isolated and shown to correspond to three nonallelic single-gene nuclear mutations. One ( noc ^r- 1 ) is semidominant while the other two ( noc ^r- 2 and noc ^r- 3 ) are recessive. Cytological and physiological studies of nocodazole's effects on the mutants demonstrate that their resistance is due neither to a lack of drug penetration nor to its degradation since, in each mutant in the presence of the drug, some microtubule networks are normal or subnormal while others remain affected as in wild-type cells. These are the first mutants resistant to microtubule depolymerizing drugs obtained in ciliates that provide a new tool for studying the assembly and dynamics of the diverse microtubule arrays in this type of organism.