title: Oligomerization of the telomerase reverse transcriptase from Euplotes crassus.
abstract: The telomerase ribonucleoprotein reverse transcriptase uses its RNA subunit as a template to synthesize telomeric repeats and maintain telomere tracts on chromosome ends. In the ciliate Euplotes crassus, the core telomerase ribonucleoprotein particle undergoes a developmentally programmed assembly into three higher order complexes after mating. Here, we provide evidence using oligonucleotide-directed affinity purification that all of the E.crassus telomerase complexes contain at least two enzyme active sites. Furthermore, we show using co-immunoprecipitation experiments that EcTERT, the telomerase catalytic subunit, undergoes multimerization in vitro. Two independent interaction domains were identified in EcTERT, one at the N-terminus that spans amino acids 186-354 and one at the C-terminus that spans amino acids 755-857. Unexpectedly, we found that TERT can form head-to-head, tail-to-tail and head-to-tail oligomers in vitro, implying that E.crassus telomerase has the potential to assume different conformations in vivo. Together, these data indicate that oligomerization is a conserved feature of telomerase and that the minimal functional unit of the enzyme is a dimer.
authors.affiliation: Department of Biochemistry and Biophysics, 2128 TAMU, Texas A&M University, College Station, TX 77843-2128, USA.
authors.forename: Libin
authors.initials: L
authors.lastname: Wang
authors.forename: Sierra R
authors.initials: SR
authors.lastname: Dean
authors.forename: Dorothy E
authors.initials: DE
authors.lastname: Shippen
mesh_terms: D000818:Animals; D001665:Binding Sites; D004268:DNA-Binding Proteins; D019281:Dimerization; D016812:Euplotes / Q000201:enzymology*; D008969:Molecular Sequence Data; D011233:Precipitin Tests; D011487:Protein Conformation; D012261:Ribonucleoproteins / Q000378:metabolism; D019098:Telomerase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D004268:DNA-Binding Proteins; D012261:Ribonucleoproteins; D019098:Telomerase