DNA double-strand breaks, but not crossovers, are required for the reorganization of meiotic nuclei in Tetrahymena.
During meiosis, the micronuclei of the ciliated protist Tetrahymena thermophila elongate dramatically. Within these elongated nuclei, chromosomes are arranged in a bouquet-like fashion and homologous pairing and recombination takes place. We studied meiotic chromosome behavior in Tetrahymena in the absence of two genes, SPO11 and a homolog of HOP2 (HOP2A), which have conserved roles in the formation of meiotic DNA double-strand breaks (DSBs) and their repair, respectively. Single-knockout mutants for each gene display only a moderate reduction in chromosome pairing, but show a complete failure to form chiasmata and exhibit chromosome missegregation. The lack of SPO11 prevents the elongation of meiotic nuclei, but it is restored by the artificial induction of DSBs. In the hop2ADelta mutant, the transient appearance of gamma-H2A.X and Rad51p signals indicates the formation and efficient repair of DSBs; but this repair does not occur by interhomolog crossing over. In the absence of HOP2A, the nuclei are elongated, meaning that DSBs but not their conversion to crossovers are required for the development of this meiosis-specific morphology. In addition, by in silico homology searches, we compiled a list of likely Tetrahymena meiotic proteins as the basis for further studies of the unusual synaptonemal complex-less meiosis in this phylogenetically remote model organism.
Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA), A-1030 Vienna, Austria.
Kazufumi
K
Mochizuki
Maria
M
Novatchkova
Josef
J
Loidl
D000595:Amino Acid Sequence; D000818:Animals; D018797:Cell Cycle Proteins / Q000235:genetics* / Q000378:metabolism; D002467:Cell Nucleus / Q000235:genetics* / Q000378:metabolism / Q000648:ultrastructure; D023902:Chromosome Pairing; D003434:Crossing Over, Genetic; D053903:DNA Breaks, Double-Stranded; D004706:Endodeoxyribonucleases; D004950:Esterases / Q000235:genetics* / Q000378:metabolism; D049469:Meiotic Prophase I / Q000235:genetics*; D008969:Molecular Sequence Data; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D013768:Tetrahymena / Q000235:genetics* / Q000378:metabolism; D000099257:Meiotic Recombination Protein SPO11
D018797:Cell Cycle Proteins; D004706:Endodeoxyribonucleases; D004950:Esterases; D029701:Saccharomyces cerevisiae Proteins; D000099257:Meiotic Recombination Protein SPO11; C113830:HOP2 protein, S cerevisiae