Figure 3.

Cdc13p binds tailed duplex molecules. Gel mobility-shift assays were carried out using 10 μg of extract from E. coli cells expressing Cdc13p or carrying vector alone. ³²P-labeled TG19 was annealed to various oligonucleotides to generate substrates with 9 bp of duplex DNA and a 10-base ss TG_1–3 tail (TG19/CAS9), with 14 bp of duplex DNA and a 5-base ss TG_1–3 tail (TG19/CAS14), or duplex TG19 (TG19/CAS19). The arrow indicates the position of the Cdc13p-DNA complex. The complex migrating more slowly than the Cdc13p-DNA complex in the vector-alone lanes was present in reduced amounts in lane 8, compared with lanes 2 and 5, and absent in lane 11 because it was due to an E. coli-encoded ss TG_1–3 DNA binding activity, and there was less (lane 8) or no (lane 11) ss TG_1–3 DNA in the DNA substrates used in those lanes.