The CCMV capsid displays quaternary structural interactions that are unique compared with previously determined RNA virus structures. The loosely coupled hexamer and pentamer morphological units readily explain their versatile reassembly properties and the pH and metal ion dependent polymorphism observed in the virions. Association of capsomeres through inter-penetrating carboxy-terminal portions of the subunit polypeptides has been previously described only for the DNA tumor viruses, SV40 and polyoma.

Motors generating mechanical force, powered by the hydrolysis of ATP, translocate doublestranded DNA into preformed capsids (proheads) of bacterial viruses 1,2 and certain animal viruses 3 . Here we describe the motor that packages the double-stranded DNA of the Bacillus subtilis bacteriophage ϕ29 into a precursor capsid. We determined the structure of the head-tail connector-the central component of the ϕ29 DNA packaging motor-to 3.2Å resolution by means of X-ray crystallography. We then fitted the connector into the electron densities of the prohead and of the partially packaged prohead as determined using cryo-electron microscopy and image reconstruction analysis. Our results suggest that the prohead plus dodecameric connector, prohead RNA, viral ATPase and DNA comprise a rotary motor with the head-prohead RNAATPase complex acting as a stator, the DNA acting as a spindle, and the connector as a ball-race. The helical nature of the DNA converts the rotary action of the connector into translation of the DNA.The bacteriophage ϕ29 (Fig. 1) is a 19-kilobase (19-kb) double-stranded DNA (dsDNA) virus with a prolate head and complex structure 4 . The prohead (Fig. 1), into which the DNA is packaged, is about 540Å long and 450Å wide 5 . The ϕ29 connector, a cone-shaped dodecamer of gene product 10 (gp10), occupies the pentagonal vertex at the base of the prohead 5 and is the portal for DNA entry during packaging and DNA ejection during infection 6 . The connector, in association with the oligomeric, ϕ29-encoded prohead RNA (pRNA) and a viral ATPase (gp16), is required for DNA packaging [7][8][9] . However, only the first 120 bases of the 174-base pRNA are essential for packaging 7 the genomic dsDNA with gp3 (DNA-gp3) can be packaged into proheads in about three minutes in vitro (P.J.J., unpublished results). The connector proteins of tailed phages 6 vary in relative molecular mass (M r ) from 36,000 (36K) in ϕ29 to 83K in phage P22, and assemble into oligomers with a central channel. The structure of the isolated ϕ29 connector has been studied by atomic force microscopy 10 and cryo-electron microscopy (cryo-EM) of two-dimensional arrays 11 , immuno-electron microscopy 12 and X-ray crystallography 13,14 .The connector structure, as now determined by X-ray crystallography, can be divided into three, approximately cylindrical regions: the narrow end, the central part, and the wide end, having external radii (Å ) of 33, 47 and 69, respectively (Fig. 2). These regions are respectively 25, 28 and 22Å in height, making the total connector 75Å long. The internal channel has a diameter of about 36Å at the narrow end, increasing to 60Å at the wide end.Comparison with electron microscopy reconstructions 5,11 shows that the narrow end protrudes from the portal vertex of the phage head, is associated with the multimeric pRNA, and binds the lower collar in the mature virus.The electron density of the connector was interpreted in terms of the amino-acid sequence 15 and was confirmed by the two Hg sites (see Methods section)...

The first structure of a flavivirus has been determined by using a combination of cryoelectron microscopy and fitting of the known structure of glycoprotein E into the electron density map. The virus core, within a lipid bilayer, has a less-ordered structure than the external, icosahedral scaffold of 90 glycoprotein E dimers. The three E monomers per icosahedral asymmetric unit do not have quasiequivalent symmetric environments. Difference maps indicate the location of the small membrane protein M relative to the overlaying scaffold of E dimers. The structure suggests that flaviviruses, and by analogy also alphaviruses, employ a fusion mechanism in which the distal beta barrels of domain II of the glycoprotein E are inserted into the cellular membrane.

Dengue virus, a member of the Flaviviridae family, has a surface composed of 180 copies each of the envelope (E) glycoprotein and the membrane (M) protein. The crystal structure of an N-terminal fragment of E has been determined and compared with a previously described structure. The primary difference between these structures is a 10 degrees rotation about a hinge relating the fusion domain DII to domains DI and DIII. These two rigid body components were used for independent fitting of E into the cryo-electron microscopy maps of both immature and mature dengue viruses. The fitted E structures in these two particles showed a difference of 27 degrees between the two components. Comparison of the E structure in its postfusion state with that in the immature and mature virions shows a rotation approximately around the same hinge. Flexibility of E is apparently a functional requirement for assembly and infection of flaviviruses.