Figure 3.
COT1 and POD6 act in the same genetic pathway. (A) A cot-1;pod-6 double mutant displayed the same morphological defects as the two parental strains. All three strains were grown on minimal media plates shifted to restrictive temperature for 5 h after growth at permissive temperature. (B) Calcofluor White stained primarily the hyphal apex of mutant strains grown at permissive temperature but mutants shifted to restrictive temperature for 5 h showed an abnormal and patchy label throughout the hyphae, including strong septal staining. (C) Suppressor analysis of cot-1(1) and pod-6(31-21) showed that gul-1 as well as components of the dynein/dynactin complex partially suppress the cot-1(1) or pod-6(31-21) growth defects (ro-1: dynein heavy chain mutant, ro-3: p150glued mutant; ro-10: 24-kDa subunit of dynactin). (D) Western-blot analysis of cell extracts probed with anti-COT1 antibody indicated that suppression of the cot-1(1) phenotype at restrictive temperature is independent of 67-kDa COT1 presence.