We have isolated about five hundred temperature-sensitive mutants specific for the mitochondrial functions. Their growth on glycerol is defective at 36 degrees C and/or 20 degrees C. While most of the mutations were nuclearly inherited, about thirty were found to be of mitochondrial origin. 1) Four mitochondrial mutations (three cryosensitive, one thermosensitive) were localized close to chloramphenicol and erythromycin resistance loci of the mitochondrial DNA, that is in the region coding for the 23 S ribosomal RNA. One of the mutation interfered with the expression of the chloramphenicol resistance gene. 2) A dozen nuclear mutations were isolated from a strain which is labelled with mitochondrial drug resistance markers (chloramphenicol, erythromycin, and paromomycin). Among the temperature sensitive respiratory deficient mutants, we have selected the mutations that supress the resistant phenotypes. We describe two non allelic such mutations, one being cryosensitive, the other thermosensitive. Both supress the expression of the mitochondrial chloramphenicol resistance gene. The temperature sensitive growth on glycerol and the modified antibiotic phenotype segregated together as a single recessive mutation. A biochemical study of these mutants is presented in a joint paper, confirming their presumed ribosomal nature.