Developmentally programmed assembly of higher order telomerase complexes with distinct biochemical and structural properties.
In Euplotes crassus, telomerase is responsible for telomere maintenance during vegetative growth and de novo telomere synthesis during macronuclear development. Here we show that telomerase in the vegetative stage of the life cycle exists as a 280-kD complex that can add telomeric repeats only onto telomeric DNA primers. Following the initiation of macronuclear development, telomerase assembles into larger complexes of 550 kD, 1600 kD, and 5 MD. In the 1600-kDa and 5-MDa complexes, telomerase is more processive than in the two smaller complexes and can add telomeres de novo onto nontelomeric 3' ends. Assembly of higher order telomerase complexes is accompanied by an extended region of RNase V1 and RNase T1 protection in the telomerase RNA subunit that is not observed with telomerase from vegetatively growing cells. The protected residues encompass a highly conserved region previously proposed to serve as a platform for formation of higher order structures. These findings provide the first direct demonstration of developmentally regulated higher order telomerase complexes with unique biochemical and structural properties.
Greene
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Department of Biochemistry and Biophysics, Texas A&M University, College Station, Texas 77843-2128, USA.
Shippen
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DE
D000818:Animals; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D016054:DNA, Protozoan / Q000235:genetics / Q000378:metabolism; D016812:Euplotes / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development*; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009690:Nucleic Acid Conformation; D016053:RNA, Protozoan / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012261:Ribonucleoproteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity; D019098:Telomerase / Q000737:chemistry* / Q000378:metabolism*; D016615:Telomere / Q000235:genetics / Q000378:metabolism
D017931:DNA Primers; D016054:DNA, Protozoan; D046911:Macromolecular Substances; D016053:RNA, Protozoan; D012261:Ribonucleoproteins; D019098:Telomerase