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Single Particle Reconstruction of the Vault from TEP1 Knockout Mice and Refinement of the Computationally Selected Vault Barrel

Published online by Cambridge University Press:  02 July 2020

P.L. Stewart ,
L.B. Kong ,
A.C. Siva ,
V.A. Kickhoefer ,
L. Harrington  and
L.H. Rome
P.L. Stewart
Affiliation:
Crump Institute for Biological Imaging and Department of Molecular & Medical Pharmacology, UCLA School of Medicine, Los Angeles, CA90095
L.B. Kong
Affiliation:
Crump Institute for Biological Imaging and Department of Molecular & Medical Pharmacology, UCLA School of Medicine, Los Angeles, CA90095
A.C. Siva
Affiliation:
Department of Biological Chemistry, UCLA School of Medicine, Los Angeles, CA90095
V.A. Kickhoefer
Affiliation:
Department of Biological Chemistry, UCLA School of Medicine, Los Angeles, CA90095
L. Harrington
Affiliation:
Department of Medical Biophysics, Ontario Cancer Institute-Amgen Institute, University of Toronto, Toronto, OntarioM5G 2C1, Canada
L.H. Rome
Affiliation:
Department of Biological Chemistry, UCLA School of Medicine, Los Angeles, CA90095
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Extract

The vault complex is a ubiquitous and predominantly cytoplasmic 13 MDa ribonucleoprotein assembly, composed of three proteins (TEP1, 240 kDa; VPARP, 193 kDa; and MVP, 100 kDa) and an untranslated RNA (vRNA) Although the basic cellular function of the vault is still unclear, recently it has been shown that induction of the major vault protein (MVP) has a direct negative influence on the nuclear uptake of the anti-cancer drug, doxorubicin. We have been applying cryo-EM single particle reconstruction methods to study the structure and molecular architecture of this cellular component. A published reconstruction of the intact rat vault at 31 Å resolution revealed that the complex is hollow and is structurally well suited to serve in macromolecular transport or sequestration. Higher resolution, 23 Å, was achieved for a reconstruction of the RNase-treated rat vault and difference imaging with the intact rat vault localized the vRNA to the ends of the vault caps.

Type
Electron Cryomicroscopy of Macromolecules
Information
Microscopy and Microanalysis , Volume 6 , Issue S2: Proceedings: Microscopy & Microanalysis 2000, Microscopy Society of America 58th Annual Meeting, Microbeam Analysis Society 34th Annual Meeting, Microscopical Society of Canada/Societe de Microscopie de Canada 27th Annual Meeting, Philadelphia, Pennsylvania August 13-17, 2000 , August 2000 , pp. 268 - 269
Copyright
Copyright © Microscopy Society of America

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References

1.Kitazono, M. et al., “Multidrug Resistance and the Lung Resistance-Related Protein in Human Colon Carcinoma SW-620 Cells” J. Natl. Cancer Inst. (1999) 91:1647.CrossRefGoogle Scholar
2.Kong, L. B. et al., “Structure of the Vault, a Ubiquitous Cellular Component.” Structure with Folding & Design (1999)7:371.CrossRefGoogle Scholar
3.Kong, L. B. et al., “RNA Localization and Modeling of a WD40 Repeat Domain within the Vault.” Submitted.Google Scholar
4. This research was supported by a grant to P.L.S. from the National Science Foundation (MCB-9722353). L.H.R. and V.A.K. acknowledge a grant from the National Institutes of Health USPHS (GM38097) and funding from the University of California (UC) BioSTAR program.Google Scholar