title: The architecture of Tetrahymena telomerase holoenzyme.
abstract: Telomerase adds telomeric repeats to chromosome ends using an internal RNA template and a specialized telomerase reverse transcriptase (TERT), thereby maintaining genome integrity. Little is known about the physical relationships among protein and RNA subunits within a biologically functional holoenzyme. Here we describe the architecture of Tetrahymena thermophila telomerase holoenzyme determined by electron microscopy. Six of the seven proteins and the TERT-binding regions of telomerase RNA (TER) have been localized by affinity labelling. Fitting with high-resolution structures reveals the organization of TERT, TER and p65 in the ribonucleoprotein (RNP) catalytic core. p50 has an unanticipated role as a hub between the RNP catalytic core, p75-p19-p45 subcomplex, and the DNA-binding Teb1. A complete in vitro holoenzyme reconstitution assigns function to these interactions in processive telomeric repeat synthesis. These studies provide the first view of the extensive network of subunit associations necessary for telomerase holoenzyme assembly and physiological function.
authors.affiliation: Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, California 90095, USA.
authors.forename: Jiansen
authors.initials: J
authors.lastname: Jiang
authors.forename: Edward J
authors.initials: EJ
authors.lastname: Miracco
authors.forename: Kyungah
authors.initials: K
authors.lastname: Hong
authors.forename: Barbara
authors.initials: B
authors.lastname: Eckert
authors.forename: Henry
authors.initials: H
authors.lastname: Chan
authors.forename: Darian D
authors.initials: DD
authors.lastname: Cash
authors.forename: Bosun
authors.initials: B
authors.lastname: Min
authors.forename: Z Hong
authors.initials: ZH
authors.lastname: Zhou
authors.forename: Kathleen
authors.initials: K
authors.lastname: Collins
authors.forename: Juli
authors.initials: J
authors.lastname: Feigon
mesh_terms: D020134:Catalytic Domain; D020035:Holoenzymes / Q000737:chemistry / Q000235:genetics / Q000648:ultrastructure; D008854:Microscopy, Electron; D008958:Models, Molecular; D009690:Nucleic Acid Conformation; D018583:Pliability; D017434:Protein Structure, Tertiary; D021122:Protein Subunits / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D015800:Protozoan Proteins / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D012313:RNA / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D012261:Ribonucleoproteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure; D019098:Telomerase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure*; D016808:Tetrahymena thermophila / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics / Q000648:ultrastructure
chemical_list: D020035:Holoenzymes; D021122:Protein Subunits; D015800:Protozoan Proteins; D012261:Ribonucleoproteins; C422952:telomerase RNA; D012313:RNA; D019098:Telomerase
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