MY01, a class XIV myosin, affects developmentally-regulated elimination of the macronucleus during conjugation of Tetrahymena thermophila.
BACKGROUND
Nuclear dimorphism is characteristic of ciliated protozoa. A transcriptionally-active macronucleus co-exists with a transcriptionally-silent micronucleus, which is activated only at conjugation. During conjugation, each conjugant develops two new genetically matched macronuclei and micronuclei, and the pre-existing macronucleus is eliminated. Elimination of the pre-existing macronucleus during conjugation is an apoptotic-like process. The macronucleus becomes highly condensed, DNA laddering occurs, caspase activity increases, acidic enzymes accumulate within the nucleoplasm, and the nucleus shrinks in size. The current study focused on the involvement of actin and myosin in nuclear events of conjugation. A myosin knockout strain was mated with wild-type, and the nuclear events were monitored with confocal microscopy.


RESULTS
Early nuclear events, including development of new macronuclei and micronuclei, appeared qualitatively normal in knockout conjugants. Completion of nuclear condensation and acidification in the pre-existing macronucleus was blocked in 44% of knockout conjugants. Knockout conjugants that failed to fully achieve nuclear condensation and acidification did not eliminate the pre-existing macronucleus. In control experiments, blockage of chromatin condensation, nuclear acidification, and macronuclear elimination was never observed in wild-type conjugants.


CONCLUSIONS
Perturbation of either DNA fragmentation, chromatin condensation or nuclear acidification can lead to blockage of apoptotic-like elimination of the macronucleus in MYO1-knockout conjugants. Consistent with the known motor function of myosins and the involvement of Myo1 in vesicle trafficking in Tetrahymena, we argue that Myo1 could specifically affect condensation of chromatin and acidification of the nucleus through direct interaction with chromatin and through Myo1-dependent vesicle trafficking to the nucleus.
Garcés
Jonathan
J
Macaulay Honors College, City University of New York, Brooklyn College, Brooklyn, NY 11210, USA.
Hosein
Roland E
RE
Gavin
Ray H
RH
D000818:Animals; D042002:Chromatin Assembly and Disassembly; D003227:Conjugation, Genetic; D048648:Macronucleus / Q000235:genetics* / Q000378:metabolism; D009218:Myosins / Q000235:genetics / Q000378:metabolism*; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism*; D016808:Tetrahymena thermophila / Q000166:cytology / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism
D015800:Protozoan Proteins; D009218:Myosins