ABSTRACT The assembly of a macromolecular structure proceeds along an ordered morphogenetic pathway, and is accomplished by the switching of proteins between discrete conformations as they

are added to the nascent assembly1,2,3. Scaffolding proteins often play a catalytic role in the assembly process1,2,4, rather like molecular chaperones5. Although macromolecular assembly

processes are fundamental to all biological systems, they have been characterized most thoroughly in viral systems, such as the icosahedral _Escherichia coli_ bacteriophage φX174 (refs 6,

7). The φX174 virion contains the proteins F, G, H and J7,8. During assembly, two scaffolding proteins B and D are required for the formation of a 108S, 360-Å-diameter procapsid from

of the external scaffold yields the mature virion. Both the F and G proteins have the eight-stranded antiparallel β-sandwich motif8,11 common to many plant and animal viruses12,13. Here we

describe the structure of a procapsid-like particle at 3.5-Å resolution, showing how the scaffolding proteins coordinate assembly of the virus by interactions with the F and G proteins, and

showing that the F protein undergoes conformational changes during capsid maturation. Access through your institution Buy or subscribe This is a preview of subscription content, access via

your institution ACCESS OPTIONS Access through your institution Subscribe to this journal Receive 51 print issues and online access $199.00 per year only $3.90 per issue Learn more Buy this

article * Purchase on SpringerLink * Instant access to full article PDF Buy now Prices may be subject to local taxes which are calculated during checkout ADDITIONAL ACCESS OPTIONS: * Log in

* Learn about institutional subscriptions * Read our FAQs * Contact customer support SIMILAR CONTENT BEING VIEWED BY OTHERS ASSEMBLY MECHANISM OF THE PLEOMORPHIC IMMATURE POXVIRUS SCAFFOLD

Article Open access 31 March 2022 CRYO-ELECTRON MICROSCOPY STRUCTURES OF CAPSIDS AND IN SITU PORTALS OF DNA-DEVOID CAPSIDS OF HUMAN CYTOMEGALOVIRUS Article Open access 11 April 2023

STRUCTURE OF THE MATURE ROUS SARCOMA VIRUS LATTICE REVEALS A ROLE FOR IP6 IN THE FORMATION OF THE CAPSID HEXAMER Article Open access 28 May 2021 REFERENCES * King, J. in _Biological

Regulation and Development_ VOL. 2, _Molecular Organization and Cell Function_(ed. Goldberger, R. F.) 101–132 (Plenum, New York, (1980). Book  Google Scholar  * Casjens, S. & Hendrix, R.

in _The Bacteriophages_ VOL. 1(ed. Calendar, R.) 15–75 (Plenum, New York, (1988). Book  Google Scholar  * Oosawa, F. & Asakura, S. _Thermodynamics of the Polymerization of

Proteins_(Academic, London, (1975). Google Scholar  * Preston, V. G., al-Kobaisi, M. F., McDougall, I. M. & Rixon, F. J. The herpes simplex virus gene UL26 proteinase in the presence of

the UL26.5 gene product promotes the formation of scaffold-like structures. _J. Gen. Virol._ 75, 2355–2366 (1994). Article  CAS  Google Scholar  * Martin, J. & Hartl, F. U.

Chaperone-assisted protein folding. _Curr. Opin. Struct. Biol._ 7, 41–52 (1997). Article  CAS  Google Scholar  * Hayashi, M. in _The Single-Stranded DNA Phages_(eds Denhardt, D. T.,

Dressler, D. & Ray, D. S.) 531–547 (Cold Spring Harbor Laboratory Press, NY, (1978). Google Scholar  * Hayashi, M., Aoyama, A., Richardson, D. L. J & Hayashi, M. N. in _The

Bacteriophages_ VOL. 2(ed. Calendar, R.) 1–71 (Plenum, New York, (1988). Book  Google Scholar  * McKenna, R. _et al_. Atomic structure of single-stranded DNA bacteriophage φX174 and its

functional implications. _Nature_ 355, 137–143 (1992). Article  ADS  CAS  Google Scholar  * Mukai, R., Hamatake, R. K. & Hayashi, M. Isolation and identification of bacteriophage φX174

prohead. _Proc. Natl Acad. Sci. USA_ 76, 4877–4881 (1979). Article  ADS  CAS  Google Scholar  * Ilag, L. L. _et al_. DNA packaging intermediates of bacteriophage φX174. _Structure_ 3,

353–363 (1995). Article  CAS  Google Scholar  * McKenna, R., Ilag, L. L. & Rossmann, M. G. Analysis of the single-stranded DNA bacteriophage φX174, refined at a resolution of 3.0 Å. _J.

Mol. Biol._ 237, 517–543 (1994). Article  CAS  Google Scholar  * Harrison, S. C., Skehel, J. J. & Wiley, D. C. in _Fields Virology_ VOL. 1(eds Fields, B. N., Knipe, D. M. & Howley,

P. M.) 59–99 (Lippincott-Raven, Philadelphia, (1996). Google Scholar  * Rossmann, M. G. & Johnson, J. E. Icosahedral RNA virus structure. _Annu. Rev. Biochem._ 58, 533–573 (1989).

Article  CAS  Google Scholar  * Rossmann, M. G. The molecular replacement method. _Acta Crystallogr. A_ 46, 73–82 (1990). Article  Google Scholar  * Dokland, T. & Murialdo, H. Structural

transitions during maturation of bacteriophage lambda capsids. _J. Mol. Biol._ 233, 682–694 (1993). Article  CAS  Google Scholar  * Prasad, B. V. V. _et al_. Three-dimensional

transformation of capsids associated with genome packaging in a bacterial virus. _J. Mol. Biol._ 231, 65–74 (1993). Article  CAS  Google Scholar  * McKenna, R., Bowman, B. R., Ilag, L. L.,

Rossmann, M. G. & Fane, B. A. Atomic structure of the degraded procapsid particle of the bacteriophage G4: induced structural changes in the presence of calcium ions and functional

implications. _J. Mol. Biol._ 256, 736–750 (1996). Article  CAS  Google Scholar  * Caspar, D. L. D. & Klug, A. Physical principles in the construction of regular viruses. _Cold Spring

Harb. Symp. Quant. Biol._ 27, 1–24 (1962). Article  CAS  Google Scholar  * Farber, M. B. Purification and properties of bacteriophage φX174 gene D products. _J. Virol._ 17, 1027–1037 (1976).

CAS  PubMed  PubMed Central  Google Scholar  * Ekechukwu, M. C. & Fane, B. A. Characterization of the morphogenetic defects conferred by cold-sensitive prohead accessory and scaffolding

proteins of φX174. _J. Bacteriol._ 177, 829–830 (1995). Article  CAS  Google Scholar  * Fane, B. A., Shien, S. & Hayashi, M. Second-site suppressors of a cold sensitive external

scaffolding protein of bacteriophage φX174. _Genetics_ 134, 1003–1011 (1993). CAS  PubMed  PubMed Central  Google Scholar  * Ekechukwu, M. C., Oberste, D. J. & Fane, B. A. Host and φX174

mutations affecting the morphogenesis or stabilization of the 50S complex, a single-stranded DNA synthesizing intermediate. _Genetics_ 140, 1167–1174 (1995). CAS  PubMed  PubMed Central 

Google Scholar  * Otwinowski, Z. & Minor, W. Processing of X-ray diffraction data collected in oscillation mode. _Methods Enzymol._ 276A, 307–326 (1997). Article  Google Scholar  *

Collaborative Computational Project Number 4. The _CCP4_ suite: programs for protein crystallography. _Acta Crystallogr. D_ 50, 760–763 (1994). Article  Google Scholar  * Tong, L. &

Rossmann, M. G. The locked rotation function. _Acta Crystallogr. A_ 46, 783–792 (1990). Article  Google Scholar  * Rossmann, M. G. _et al_. Molecular replacement real-space averaging. _J.

Appl. Crystallogr._ 25, 166–180 (1992). Article  CAS  Google Scholar  * Jones, T. A., Zou, J. Y., Cowan, S. W. & Kjeldgaard, M. Improved methods for building protein models in electron

density maps and the location of errors in these models. _Acta Crystallogr. A_ 47, 110–119 (1991). Article  Google Scholar  * Rossmann, M. G. & Blow, D. M. The detection of sub-units

within the crystallographic asymmetric unit. _Acta Crystallogr._ 15, 24–31 (1962). Article  CAS  Google Scholar  * Kraulis, P. MOLSCRIPT: a program to produce both detailed and schematic

plots of protein structures. _J. Appl. Crystallogr._ 24, 946–950 (1991). Article  Google Scholar  Download references ACKNOWLEDGEMENTS We thank the staff at the Cornell High Energy

Synchrotron Source and the National Light Source staff at beam line X12 for providing the data collection facilities; W. F. Bean for help with computational work; and S. Wilder for help in

the preparation of this manuscript. This study was supported by a National Science Foundation grant to M.G.R., an NIH grant to B.A.F., a Robert and Richard Rizzo Memorial Fund grant and an

NIH grant to N.L.I., a Lucille P. Markey Foundation award to Purdue University, and a Purdue University reinvestment program grant. T.D. was supported by a European Molecular Biology

Organization long-term postdoctoral fellowship for some of the time of this work. AUTHOR INFORMATION AUTHORS AND AFFILIATIONS * Department of Biological Sciences, Purdue University, West

Lafayette, 47907-1392, Indiana, USA Terje Dokland, Robert McKenna, Leodevico L. Ilag, Brian R. Bowman & Michael G. Rossmann * Department of Biological Sciences, Warwick University, CV4

7AL, Coventry, UK Robert McKenna, Leodevico L. Ilag & Brian R. Bowman * Morphosys GmbH, Frankfurter Ring 193a, 80807, München, Germany Robert McKenna, Leodevico L. Ilag & Brian R.

Bowman * Department of Biochemistry, Baylor College of Medicine, Houston, 77030, Texas, USA Robert McKenna, Leodevico L. Ilag & Brian R. Bowman * Department of Microbiology and

Immunology, University of Tennessee, Memphis, 38163, Tennessee, USA Nino L. Incardona * Department of Biological Sciences, University of Arkansas, Fayetteville, 72701, Arkansas, USA Bentley

A. Fane Authors * Terje Dokland View author publications You can also search for this author inPubMed Google Scholar * Robert McKenna View author publications You can also search for this

author inPubMed Google Scholar * Leodevico L. Ilag View author publications You can also search for this author inPubMed Google Scholar * Brian R. Bowman View author publications You can

also search for this author inPubMed Google Scholar * Nino L. Incardona View author publications You can also search for this author inPubMed Google Scholar * Bentley A. Fane View author

publications You can also search for this author inPubMed Google Scholar * Michael G. Rossmann View author publications You can also search for this author inPubMed Google Scholar RIGHTS AND

PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Dokland, T., McKenna, R., Ilag, L. _et al._ Structure of a viral procapsid with molecular scaffolding. _Nature_

389, 308–313 (1997). https://doi.org/10.1038/38537 Download citation * Received: 24 March 1997 * Accepted: 10 June 1997 * Issue Date: 18 September 1997 * DOI: https://doi.org/10.1038/38537

SHARE THIS ARTICLE Anyone you share the following link with will be able to read this content: Get shareable link Sorry, a shareable link is not currently available for this article. Copy to

clipboard Provided by the Springer Nature SharedIt content-sharing initiative