The yeast centromere CDEI/Cpf1 complex: differences between in vitro binding and in vivo function

摘要: The centromere and promoter factor Cpf1 binds DNA element I found in all DNAs from the yeast Saccharomyces cerevisiae. We analyzed thirty different point mutations or around CEN6-CDEI (ATCACGTG) for their relative binding affinity to these data were compared with vivo function of mutants. show that minimal length site needed full vitro activity is 10 base pairs long comprised CDEI plus two 3' this sequence. palindromic core sequence CACGTG most important CEN binding. Symmetrical either halfsite affect mitotic asymmetrically albeit a extent. Enlarging palindrome 12 20 bps increases but results increased chromosome loss rates suggesting need asymmetrical sequences. Additionally, ability protein bind mutant does not parallel confer activity. Our indicate characteristics only partly overlap corresponding within complex, thus situation CDEI/Cpf1 complex might undergo interactions other DNA/protein complexes.