Caspase-like activity is required for programmed nuclear elimination during conjugation in Tetrahymena.
During conjugation in the binucleate ciliate, Tetrahymena thermophila, the old macronucleus is eliminated as new macronuclei and micronuclei are ontogenetically derived from the zygote nucleus. The mechanism of programmed nuclear elimination in ciliates may be related to the mechanism of apoptosis in higher organisms since its chromatin undergoes major condensation, its DNA is digested into nucleosome-sized fragments, and it stains positively for TUNEL. The present study explores whether caspases are involved in programmed macronuclear degradation in Tetrahymena. We show here that caspase-like activity is detectable using two specific colorimetric substrates, and that the activity is reduced with specific caspase inhibitors. In addition, using the fluorigenic substrate PhiPhiLux, active caspase-like activity is detected in living cells, localized to cytoplasmic vesicles; activity is not detected in pre- or post-condensed macronuclei. Finally, three different inhibitors of caspase activity cause a block to macronuclear chromatin condensation and elimination. Therefore, a caspase-like enzyme activity is necessary for regulating macronuclear elimination in Tetrahymena. These data support the possibility that macronuclear elimination is related, evolutionarily, to regulated cell death in multicellular organisms.
Ejercito
Mylee
M
Biology Department, Wesleyan University, Middletown, CT 06459, USA.
Wolfe
Jason
J
D000818:Animals; D020169:Caspases / Q000378:metabolism*; D002467:Cell Nucleus / Q000648:ultrastructure*; D002843:Chromatin / Q000235:genetics / Q000648:ultrastructure*; D003227:Conjugation, Genetic / Q000235:genetics*; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D007700:Kinetics; D016808:Tetrahymena thermophila / Q000235:genetics* / Q000648:ultrastructure*
D002843:Chromatin; D015853:Cysteine Proteinase Inhibitors; D020169:Caspases