The Saccharomyces telomere-binding protein Cdc13p interacts with both the catalytic subunit of DNA polymerase alpha and the telomerase-associated est1 protein.
Saccharomyces telomeres consist of approximately 350 bp of C(1-3)A/TG(1-3) DNA. Most of this approximately 350 bp is replicated by standard, semiconservative DNA replication. After conventional replication, the C(1-3)A strand is degraded to generate a long single strand TG(1-3) tail that can serve as a substrate for telomerase. Cdc13p is a single strand TG(1-3) DNA-binding protein that localizes to telomeres in vivo. Genetic data suggest that the Cdc13p has multiple roles in telomere replication. We used two hybrid analysis to demonstrate that Cdc13p interacted with both the catalytic subunit of DNA polymerase alpha, Pol1p, and the telomerase RNA-associated protein, Est1p. The association of these proteins was confirmed by biochemical analysis using full-length or nearly full-length proteins. Point mutations in either CDC13 or POL1 that reduced the Cdc13p-Pol1p interaction resulted in telomerase mediated telomere lengthening. Over-expression of the carboxyl terminus of Est1p partially suppressed the temperature sensitive lethality of a cdc13-1 strain. We propose that Cdc13p's interaction with Est1p promotes TG(1-3) strand lengthening by telomerase and its interaction with Pol1p promotes C(1-3)A strand resynthesis by DNA polymerase alpha.
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Department of Molecular Biology, Princeton University, Princeton, New Jersey 08544-1014, USA.
Zakian
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D000483:Alleles; D002384:Catalysis; D019926:Cyclin B / Q000737:chemistry / Q000378:metabolism*; D004256:DNA Polymerase I / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005656:Fungal Proteins / Q000737:chemistry / Q000378:metabolism*; D005982:Glutathione Transferase / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D010957:Plasmids; D017354:Point Mutation; D011485:Protein Binding; D012440:Saccharomyces / Q000737:chemistry* / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins; D019098:Telomerase / Q000737:chemistry / Q000378:metabolism*; D016615:Telomere / Q000235:genetics / Q000378:metabolism; D020798:Two-Hybrid System Techniques
D019926:Cyclin B; D005656:Fungal Proteins; D029701:Saccharomyces cerevisiae Proteins; D005982:Glutathione Transferase; C099024:EST1 protein, S cerevisiae; D019098:Telomerase; D004256:DNA Polymerase I; C083536:POL1 protein, S cerevisiae