|dc.description.abstract||Telomerase is the key enzyme synthesizing telomeric DNA in most eukaryotic organisms. In mammals, telomerase expression is abundant in the germline cells but is undetectable in most other differentiated organs. Intensive studies of telomerase have focused on human cancerous cells, where over 90% of all cancerous tissues examined have telomerase activity. In wild-type Arabidopsis, telomerase expression is abundant in reproductive organs and dedifferentiated tissues such as flowers, siliques and calli but barely detectable in vegetative tissues (both rosette and cauline leaves). In this study, a biochemical screen strategy was developed for isolation of telomerase activating mutants in Arabidopsis thaliana. Through screening of Arabidopsis activation-tagged lines by a PCR-based TRAP assay, two tac (for telomerase activator) mutants were isolated. RT-PCR analysis of AtTERT expression revealed that different mechanisms are involved in alternating telomerase activity in tac1 and tac2.
We cloned and characterized the TAC1 gene. TAC1 encodes a single zinc finger protein and acts synergistically with auxin to induce telomerase expression without altering cell cycles. Telomere length was unperturbed in the mutant, but other phenotypes, such as altered root development and the ability of cells to grow in culture without exogenous auxin, indicated that TAC1 not only is part of the previously reported link between auxin and telomerase expression, but also potentiates other classic responses to this phytohormone.
DNA microarrays were used to analyze the expression profile of the tac1 mutant and revealed that several drought-induced genes were up-regulated 3 to 10 fold in the tac1-1D mutant. RT-PCR analysis further confirmed this up-regulation for five of these genes. Investigation of root growth also indicated that tac1-1D roots were ~20% longer relative to wild-type. Further experiments demonstrated that over-expression of TAC1 does confer drought tolerance, but not salt tolerance. In addition, our preliminary result showed that treatment with a low concentration of IAA could induce drought tolerance in wild-type Arabidopsis. Although plants with constitutive expression of telomerase have no practical utility, the ability of TAC1 to confer drought tolerance could have significant agricultural applications.||en