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Figure 1


Fig. 1. m3G cap formation of TLC1 is dependent on Tgs1. (A) Whole cell extracts were prepared from wild-type and tgs1{Delta} cells. After precipitation using an antibody against the m3G structure, RNA was extracted and analyzed by northern blotting. For hybridization, the random primed [{alpha} 32P]dCTP-labeled TLC1 DNA was used as a probe. Input (10%), anti-m3G immunoprecipitates (RNA immunoprecipitates of the m3G antibody; 90%) and supernatant (unbound RNA; 25%) are shown. Duplicates of immunoprecipitates and supernatant were loaded. (B) Telomeres are elongated in the absence of Tgs1. Genomic DNA was prepared from wild-type and tgs1{Delta} cells and digested with XhoI. DNA fragments were analyzed by Southern blotting. As a probe, a CA-rich fragment of telomere VII was labeled with [{alpha} 32P]dCTP. Two wild-type and three tgs1{Delta} genomic DNA preparations are shown. Black and white arrows indicate the average telomere length in the WT and tgs1 mutant, respectively. (C) Telomeres are elongated in the catalytically inactive tgs1-W178A mutant. Genomic DNA was analyzed as in B. Black and white arrows indicate the average telomere length in the WT and tgs1 mutant, respectively.





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