Selfing mutants link Ku proteins to mating type determination in Tetrahymena.

Recognition of self and nonself is important for outcrossing organisms, and different mating types establish the barrier against self-mating. In the unicellular ciliate T. thermophila, mating type determination requires complex DNA rearrangements at a single mat locus during conjugation to produce a type-specific gene pair (MTA and MTB) for 1 of 7 possible mating types. Surprisingly, we found that decreased expression of the DNA breakage-repair protein Ku80 at late stages of conjugation generated persistent selfing phenotype in the progeny. DNA analysis revealed multiple mating-type gene pairs as well as a variety of mis-paired, unusually arranged mating-type genes in these selfers that resemble some proposed rearrangement intermediates. They are found also in normal cells during conjugation and are lost after 10 fissions but are retained in Ku mutants. Silencing of TKU80 or TKU70-2 immediately after conjugation also generated selfing phenotype, revealing a hidden DNA rearrangement process beyond conjugation. Mating reactions between the mutant and normal cells suggest a 2-component system for self–nonself-recognition through MTA and MTB genes. In the unicellular ciliate Tetrahymena thermophila, mating type determination requires complex DNA rearrangements to produce a type-specific MAT gene pair for each of seven possible mating types. This study shows that DNA recombination produces a large number of unusual MAT gene pairs, but through a KU-dependent process only one normal pair is retained in a mature cell; retention of unusual gene pairs leads to self-mating and suggests a system for self/non-self recognition. [ABSTRACT FROM AUTHOR]