Cryo-EM structure of a transcribing cypovirus.

Double-stranded RNA viruses in the family Reoviridae are capable of transcribing and capping nascent mRNA within an icosahedral viral capsid that remains intact throughout repeated transcription cycles. However, how the highly coordinated mRNA transcription and capping process is facilitated by viral capsid proteins is still unknown. Cypovirus provides a good model system for studying the mRNA transcription and capping mechanism of viruses in the family Reoviridae. Here, we report a full backbone model of a transcribing cypovirus built from a near-atomic-resolution density map by cryoelectron microscopy. Compared with the structure of a nontranscribing cypovirus, the major capsid proteins of transcribing cypovirus undergo a series of conformational changes, giving rise to structural changes in the capsid shell: (i) an enlarged capsid chamber, which provides genomic RNA with more flexibility to move within the densely packed capsid, and (ii) a widened peripentonal channel in the capsid shell, which we confirmed to be a pathway for nascent mRNA. A rod-like structure attributable to a partially resolved nascent mRNA was observed in this channel. In addition, conformational change in the turret protein results in a relatively open turret at each fivefold axis. A GMP moiety, which is transferred to 5'-diphosphorylated mRNA during the mRNA capping reaction, was identified in the pocket-like guanylyltransferase domain of the turret protein.

A Multiphysics Model for Predicting Microstructure Changes and Microhardness of Machined AerMet100 Steel.

The machined-surface integrity plays a critical role in corrosion resistance and fatigue properties of ultra-high-strength steels. This work develops a multiphysics model for predicting the microstructure changes and microhardness of machined AerMet100 steel. The variations of stress, strain and temperature of the machined workpiece are evaluated by constructing a finite-element model of the orthogonal cutting process. Based on the multiphysics fields, the analytical models of phase transformation and dislocation density evolution are built up. The white layer is modeled according to the phase-transformation mechanism and the effects of stress and plastic strain on real phase-transformation temperature are taken into consideration. The microhardness changes are predicted by a model that accounts for both dislocation density and phase-transformation evolution processes. Experimental tests are carried out for model validation. The predicted results of cutting force, white-layer thickness and microhardness are in good agreement with the measured data. Additionally, from the proposed model, the correlation between the machined-surface characteristics and processing parameters is established.

Effects of Multi-Pass Turning on Stress Corrosion Cracking of AISI 304 Austenitic Stainless Steel.

Austenitic stainless steels are extensively used in mechanical engineering. The machined surface integrity has an essential influence on the stress corrosion cracking (SCC) performance of stainless steels. In this paper, the effects of multi-pass turning on the SCC susceptibility of AISI 304 austenitic stainless steel were investigated by correlating the SCC crack density to the machining-induced surface characteristics in terms of roughness, micro-hardness, and residual stress. In the multi-pass turning, the surface roughness and residual stress were the least after the double pass turning, and the surface micro-hardness was the maximum after the triple-pass turning. The SCC susceptibility was evaluated after SCC tests in boiling MgCl2 solution. The results showed that the weakest SCC sensitivity was observed in double-pass turning 304 stainless steel, while the most susceptible SCC was found in triple-pass turning. Compared with the double-pass turning, the increase in SCC sensitivity of triple-pass turning was attributed to the larger roughness, higher micro-hardness and greater residual tensile stresses.

Identification of the active sites in the methyltransferases of a transcribing dsRNA virus.

Many double-stranded RNA (dsRNA) viruses are capable of transcribing and capping RNA within a stable icosahedral viral capsid. The turret of turreted dsRNA viruses belonging to the family Reoviridae is formed by five copies of the turret protein, which contains domains with both 7-N-methyltransferase and 2'-O-methyltransferase activities, and serves to catalyze the methylation reactions during RNA capping. Cypovirus of the family Reoviridae provides a good model system for studying the methylation reactions in dsRNA viruses. Here, we present the structure of a transcribing cypovirus to a resolution of ~3.8Å by cryo-electron microscopy. The binding sites for both S-adenosyl-L-methionine and RNA in the two methyltransferases of the turret were identified. Structural analysis of the turret in complex with RNA revealed a pathway through which the RNA molecule reaches the active sites of the two methyltransferases before it is released into the cytoplasm. The pathway shows that RNA capping reactions occur in the active sites of different turret protein monomers, suggesting that RNA capping requires concerted efforts by at least three turret protein monomers. Thus, the turret structure provides novel insights into the precise mechanisms of RNA methylation.

Cryo-EM structures of two bovine adenovirus type 3 intermediates.

Adenoviruses (Ads) infect hosts from all vertebrate species and have been investigated as vaccine vectors. We report here near-atomic structures of two bovine Ad type 3 (BAd3) intermediates obtained by cryo-electron microscopy. A comparison between the two intermediate structures reveals that the differences are localized in the fivefold vertex region, while their facet structures are identical. The overall facet structure of BAd3 exhibits a similar structure to human Ads; however, BAd3 protein IX has a unique conformation. Mass spectrometry and cryo-electron tomography analyses indicate that one intermediate structure represents the stage during DNA encapsidation, whilst the other intermediate structure represents a later stage. These results also suggest that cleavage of precursor protein VI occurs during, rather than after, the DNA encapsidation process. Overall, our results provide insights into the mechanism of Ad assembly, and allow the first structural comparison between human and nonhuman Ads at backbone level.