This project explores the synthesis of heterogeneous G1-3T yeast telomere sequences by the enzyme telomerase. I demonstrate that a mutant in the essential N-terminal TEN domain of Est2p (telomerase reverse transciptase), glutamic acid 76 to lysine (est2-LTE76K), restricted possible alignments between the DNA primer and the RNA template and increased in vivo processivity. Within the context of telomerase, the Est2p TEN domain is thought to contribute to enzyme processivity by mediating an anchor-site interaction with the DNA primer. I showed that binding of the purified TEN domain (residues 1-161) to telomeric DNA is enhanced by the E76K mutation. These results suggest a novel role for the anchor-site in mediating primer/template alignment within the active site of yeast telomerase.
| Identifer | oai:union.ndltd.org:VANDERBILT/oai:VANDERBILTETD:etd-08242011-132833 |
| Date | 02 September 2011 |
| Creators | Bairley, Robin Christine Brooks |
| Contributors | David Cortez, Kathy Gould, Carl Johnson, Todd Graham, Katherine Friedman |
| Publisher | VANDERBILT |
| Source Sets | Vanderbilt University Theses |
| Language | English |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | http://etd.library.vanderbilt.edu/available/etd-08242011-132833/ |
| Rights | unrestricted, I hereby certify that, if appropriate, I have obtained and attached hereto a written permission statement from the owner(s) of each third party copyrighted matter to be included in my thesis, dissertation, or project report, allowing distribution as specified below. I certify that the version I submitted is the same as that approved by my advisory committee. I hereby grant to Vanderbilt University or its agents the non-exclusive license to archive and make accessible, under the conditions specified below, my thesis, dissertation, or project report in whole or in part in all forms of media, now or hereafter known. I retain all other ownership rights to the copyright of the thesis, dissertation or project report. I also retain the right to use in future works (such as articles or books) all or part of this thesis, dissertation, or project report. |
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