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Oligomerization of the telomerase reverse transcriptase from Euplotes crassus
Libin Wang1, Sierra R Dean, Dorothy E Shippen
1Department of Biochemistry and Biophysics, 2128 TAMU, Texas A&M University, College Station, TX 77843-2128, USA.
Nucleic Acids Research
|September 18, 2002
Summary
Telomerase enzyme active sites were studied in Euplotes crassus. The telomerase catalytic subunit (EcTERT) multimerizes, forming dimers as the minimal functional unit.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- Telomerase is a ribonucleoprotein reverse transcriptase essential for maintaining telomere length.
- In Euplotes crassus, telomerase complexes assemble into higher-order structures post-mating.
Purpose of the Study:
- To investigate the active sites and oligomerization of Euplotes crassus telomerase.
- To identify interaction domains within the telomerase catalytic subunit (EcTERT).
Main Methods:
- Oligonucleotide-directed affinity purification to assess enzyme active sites.
- Co-immunoprecipitation experiments to study EcTERT multimerization in vitro.
Main Results:
- All Euplotes crassus telomerase complexes possess at least two active enzyme sites.
- EcTERT multimerizes in vitro, with identified N-terminal and C-terminal interaction domains.
- TERT forms various oligomeric structures (head-to-head, tail-to-tail, head-to-tail) in vitro.
Conclusions:
- Telomerase oligomerization is a conserved biological process.
- The minimal functional unit of Euplotes crassus telomerase is a dimer.