Jiansen Jiang / Edward J Miracco / Kyungah Hong / Barbara Eckert / Henry Chan / Darian D Cash / Bosun Min / Z Hong Zhou / Kathleen Collins / Juli Feigon /
PubMed 要旨
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 ...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.
EMDB-5804: Three-dimensional structure of endogenously assembled Tetrahymena telomerase holoenzyme determined by negative-stain electron microscopy 手法: EM (単粒子) / 解像度: 25.0 Å
EMDB-5807: Negative-stain electron microscopy reconstruction of Tetrahymena telomerase (Teb1-f, conformation II) 手法: EM (単粒子) / 解像度: 30.0 Å
EMDB-5808: Negative-stain electron microscopy reconstruction of Tetrahymena telomerase (Teb1-f, conformation III) 手法: EM (単粒子) / 解像度: 30.0 Å
EMDB-5809: Negative-stain electron microscopy reconstruction of Tetrahymena telomerase (Teb1-f, conformation IV) 手法: EM (単粒子) / 解像度: 30.0 Å
EMDB-5810: Negative-stain electron microscopy reconstruction of Tetrahymena telomerase (Teb1-f, conformation V) 手法: EM (単粒子) / 解像度: 30.0 Å
EMDB-5811: Negative-stain electron microscopy reconstruction of Tetrahymena telomerase (Teb1-f, conformation VI) 手法: EM (単粒子) / 解像度: 30.0 Å
EMDB-5812: Negative-stain electron microscopy reconstruction of Tetrahymena telomerase (Teb1-f, conformation VII) 手法: EM (単粒子) / 解像度: 30.0 Å
EMDB-5813: Negative-stain electron microscopy reconstruction of Tetrahymena telomerase (Teb1-f, conformation VIII) 手法: EM (単粒子) / 解像度: 30.0 Å
EMDB-5814: Negative-stain electron microscopy reconstruction of Tetrahymena telomerase (Teb1-f, conformation IX) 手法: EM (単粒子) / 解像度: 30.0 Å
EMDB-5815: Negative-stain electron microscopy reconstruction of Tetrahymena telomerase (Teb1-f, conformation X) 手法: EM (単粒子) / 解像度: 30.0 Å
EMDB-5816: Negative-stain electron microscopy reconstruction of Tetrahymena telomerase (TERT-f, stable conformation) 手法: EM (単粒子) / 解像度: 35.0 Å
EMDB-5817: Negative-stain electron microscopy reconstruction of Tetrahymena telomerase (TERT-f, subcomplex lacking p65 N-terminus) 手法: EM (単粒子) / 解像度: 35.0 Å
EMDB-5818: Negative-stain electron microscopy reconstruction of Tetrahymena telomerase (TERT-f, subcomplex lacking Teb1) 手法: EM (単粒子) / 解像度: 35.0 Å
EMDB-5819: Negative-stain electron microscopy reconstruction of Tetrahymena telomerase (TERT-f, subcomplex lacking p75-p19-p45) 手法: EM (単粒子) / 解像度: 35.0 Å
EMDB-5820: Negative-stain electron microscopy reconstruction of Tetrahymena telomerase (f-Teb1C, stable conformation) 手法: EM (単粒子) / 解像度: 30.0 Å
EMDB-5821: Negative-stain electron microscopy reconstruction of Tetrahymena telomerase (TERT-f, affinity labeled by anti-FLAG Fab) 手法: EM (単粒子) / 解像度: 30.0 Å
EMDB-5822: Negative-stain electron microscopy reconstruction of Tetrahymena telomerase (Teb1-f, affinity labeled by anti-FLAG Fab) 手法: EM (単粒子) / 解像度: 30.0 Å
EMDB-5823: Negative-stain electron microscopy reconstruction of Tetrahymena telomerase (f-p50, affinity labeled by two copies of anti-FLAG Fab) 手法: EM (単粒子) / 解像度: 30.0 Å
EMDB-5824: Negative-stain electron microscopy reconstruction of Tetrahymena telomerase (f-p50, affinity labeled by one copy of anti-FLAG Fab) 手法: EM (単粒子) / 解像度: 30.0 Å