Structural Basis for Telomerase RNA Recognition and RNP Assembly by the Holoenzyme La Family Protein p65

Mahavir Singh; Zhonghua Wang; Bon Kyung Koo; Anooj Patel; Duilio Cascio; Kathleen Collins; Juli Feigon

doi:10.1016/j.molcel.2012.05.018

Structural Basis for Telomerase RNA Recognition and RNP Assembly by the Holoenzyme La Family Protein p65

Mahavir Singh, Zhonghua Wang, Bon Kyung Koo, Anooj Patel, Duilio Cascio, Kathleen Collins, Juli Feigon

Research output: Contribution to journal › Article › peer-review

69 Scopus citations

Abstract

Telomerase is a ribonucleoprotein complex essential for maintenance of telomere DNA at linear chromosome ends. The catalytic core of Tetrahymena telomerase comprises a ternary complex of telomerase RNA (TER), telomerase reverse transcriptase (TERT), and the essential La family protein p65. NMR and crystal structures of p65 C-terminal domain and its complex with stem IV of TER reveal that RNA recognition is achieved by a combination of single- and double-stranded RNA binding, which induces a 105° bend in TER. The domain is a cryptic, atypical RNA recognition motif with a disordered C-terminal extension that forms an α helix in the complex necessary for hierarchical assembly of TERT with p65-TER. This work provides the first structural insight into biogenesis and assembly of TER with a telomerase-specific protein. Additionally, our studies define a structurally homologous domain (xRRM) in genuine La and LARP7 proteins and suggest a general mode of RNA binding for biogenesis of their diverse RNA targets.

Original language	English (US)
Pages (from-to)	16-26
Number of pages	11
Journal	Molecular Cell
Volume	47
Issue number	1
DOIs	https://doi.org/10.1016/j.molcel.2012.05.018
State	Published - Jul 13 2012
Externally published	Yes

ASJC Scopus subject areas

Molecular Biology
Cell Biology

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10.1016/j.molcel.2012.05.018

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title = "Structural Basis for Telomerase RNA Recognition and RNP Assembly by the Holoenzyme La Family Protein p65",

abstract = "Telomerase is a ribonucleoprotein complex essential for maintenance of telomere DNA at linear chromosome ends. The catalytic core of Tetrahymena telomerase comprises a ternary complex of telomerase RNA (TER), telomerase reverse transcriptase (TERT), and the essential La family protein p65. NMR and crystal structures of p65 C-terminal domain and its complex with stem IV of TER reveal that RNA recognition is achieved by a combination of single- and double-stranded RNA binding, which induces a 105° bend in TER. The domain is a cryptic, atypical RNA recognition motif with a disordered C-terminal extension that forms an α helix in the complex necessary for hierarchical assembly of TERT with p65-TER. This work provides the first structural insight into biogenesis and assembly of TER with a telomerase-specific protein. Additionally, our studies define a structurally homologous domain (xRRM) in genuine La and LARP7 proteins and suggest a general mode of RNA binding for biogenesis of their diverse RNA targets.",

author = "Mahavir Singh and Zhonghua Wang and Koo, {Bon Kyung} and Anooj Patel and Duilio Cascio and Kathleen Collins and Juli Feigon",

note = "Funding Information: This work was supported by NSF and NIH grants to J.F. and by NIH grant to K.C. The authors thank Drs. C.J. Park, R. Peterson, and Q. Zhang for help with NMR; K. Stahmer for help in the early stages of this work; M. Zhao for help with X-ray data collection; and the staff members of beamlines ID-24 (NE-CAT) and ID-23 (GM/CA) at APS of Argonne National Laboratory and CCP4 APS School 2011 for help with data collection and analysis. M.S. designed experiments, prepared samples, performed experiments, analyzed the data, and wrote the paper; Z.W. helped refine the NMR structure and performed sequence alignment; B.-K.K. helped with NMR structure calculations; A.P. prepared protein and RNA samples; D.C. helped with X-ray data collection and analysis; K.C. provided reagents and edited the paper; and J.F. designed experiments, analyzed data, supervised all aspects of the work, and wrote the paper. ",

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AU - Singh, Mahavir

AU - Wang, Zhonghua

AU - Koo, Bon Kyung

AU - Patel, Anooj

AU - Cascio, Duilio

AU - Collins, Kathleen

AU - Feigon, Juli

N1 - Funding Information: This work was supported by NSF and NIH grants to J.F. and by NIH grant to K.C. The authors thank Drs. C.J. Park, R. Peterson, and Q. Zhang for help with NMR; K. Stahmer for help in the early stages of this work; M. Zhao for help with X-ray data collection; and the staff members of beamlines ID-24 (NE-CAT) and ID-23 (GM/CA) at APS of Argonne National Laboratory and CCP4 APS School 2011 for help with data collection and analysis. M.S. designed experiments, prepared samples, performed experiments, analyzed the data, and wrote the paper; Z.W. helped refine the NMR structure and performed sequence alignment; B.-K.K. helped with NMR structure calculations; A.P. prepared protein and RNA samples; D.C. helped with X-ray data collection and analysis; K.C. provided reagents and edited the paper; and J.F. designed experiments, analyzed data, supervised all aspects of the work, and wrote the paper.

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N2 - Telomerase is a ribonucleoprotein complex essential for maintenance of telomere DNA at linear chromosome ends. The catalytic core of Tetrahymena telomerase comprises a ternary complex of telomerase RNA (TER), telomerase reverse transcriptase (TERT), and the essential La family protein p65. NMR and crystal structures of p65 C-terminal domain and its complex with stem IV of TER reveal that RNA recognition is achieved by a combination of single- and double-stranded RNA binding, which induces a 105° bend in TER. The domain is a cryptic, atypical RNA recognition motif with a disordered C-terminal extension that forms an α helix in the complex necessary for hierarchical assembly of TERT with p65-TER. This work provides the first structural insight into biogenesis and assembly of TER with a telomerase-specific protein. Additionally, our studies define a structurally homologous domain (xRRM) in genuine La and LARP7 proteins and suggest a general mode of RNA binding for biogenesis of their diverse RNA targets.

AB - Telomerase is a ribonucleoprotein complex essential for maintenance of telomere DNA at linear chromosome ends. The catalytic core of Tetrahymena telomerase comprises a ternary complex of telomerase RNA (TER), telomerase reverse transcriptase (TERT), and the essential La family protein p65. NMR and crystal structures of p65 C-terminal domain and its complex with stem IV of TER reveal that RNA recognition is achieved by a combination of single- and double-stranded RNA binding, which induces a 105° bend in TER. The domain is a cryptic, atypical RNA recognition motif with a disordered C-terminal extension that forms an α helix in the complex necessary for hierarchical assembly of TERT with p65-TER. This work provides the first structural insight into biogenesis and assembly of TER with a telomerase-specific protein. Additionally, our studies define a structurally homologous domain (xRRM) in genuine La and LARP7 proteins and suggest a general mode of RNA binding for biogenesis of their diverse RNA targets.

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Structural Basis for Telomerase RNA Recognition and RNP Assembly by the Holoenzyme La Family Protein p65

Abstract

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