Posttargeting Events in Cotranslational Translocation Through the Sec61 Complex: a Thesis

The cytoplasmic surface of Sec61p is the binding site for the ribosome and has been proposed to interact with the signal recognition particle receptor during targeting of the ribosome nascent chain complex to the translocation channel. Point mutations in cytoplasmic loops six (L6) and eight (L8) of yeast Sec61p cause reductions in growth rates and defects in translocation of nascent polypeptides that utilize the cotranslational translocation pathway. Sec61 heterotrimers isolated from the L8 sec61 mutants have a greatly reduced affinity for 80S ribosomes. Cytoplasmic accumulation of protein precursors demonstrates that the initial contact between the large ribosomal subunit and the Sec61 complex is important for efficient insertion of a nascent polypeptide into the translocation pore. In contrast, point mutations in L6 of Sec61p inhibit cotranslational translocation without significantly reducing the ribosome binding activity, indicating that the L6 and L8 sec61 mutants impact different steps in the cotranslational translocation pathway. Integral membrane proteins are cotranslationally inserted into the endoplasmic reticulum via the protein translocation channel, which mediates the translocation of lumenal domains, retention of cytosolic domains and integration of transmembrane spans into the phospholipid bilayer. We analyzed the in vivo kinetics of integration of model membrane proteins in Saccharomyces cerevisiae using ubiquitin translocation assay reporters. A signal anchor sequence from a type II membrane protein gates the translocon pore less rapidly than a cleavable signal sequence from a secretory protein. Transmembrane spans and lumenal domains are exposed to the cytosol during integration of a poly topic membrane protein. The conformational changes in the translocon that permit opening of the lumenal and lateral channel gates occur less rapidly than elongation of the nascent polypeptide. Cytosolic exposure of transmembrane spans and lumenal domains poses a challenge to the fidelity of membrane protein integration

An interaction between the SRP receptor and the translocon is critical during cotranslational protein translocation

The signal recognition particle (SRP)-dependent targeting pathway facilitates rapid, efficient delivery of the ribosome-nascent chain complex (RNC) to the protein translocation channel. We test whether the SRP receptor (SR) locates a vacant protein translocation channel by interacting with the yeast Sec61 and Ssh1 translocons. Surprisingly, the slow growth and cotranslational translocation defects caused by deletion of the transmembrane (TM) span of yeast SRbeta (SRbeta-DeltaTM) are exaggerated when the SSH1 gene is disrupted. Disruption of the SBH2 gene, which encodes the beta subunit of the Ssh1p complex, likewise causes a growth defect when combined with SRbeta-DeltaTM. Cotranslational translocation defects in the ssh1DeltaSRbeta-DeltaTM mutant are explained by slow and inefficient in vivo gating of translocons by RNCs. A critical function for translocation channel beta subunits in the SR-channel interaction is supported by the observation that simultaneous deletion of Sbh1p and Sbh2p causes a defect in the cotranslational targeting pathway that is similar to the translocation defect caused by deletion of either subunit of the SR

Numerical simulation of separation shock characteristics of a piston type explosive bolt

A piston type explosive bolt is modeled by using a hydrocodes AUTODYN. The influence of the charge amount on the separation shock is analyzed. The results show that the separation shock of the piston type explosive bolt mainly includes two aspects: the shock caused by explosive detonation and the impact of the piston at the end of stroke. As the charge amount increases, the collision speed of piston first increases and then decreases, and the separation shock first increases and then stabilizes

A Multilayer Perceptron-based Fast Sunlight Assessment for the Conceptual Design of Residential Neighborhoods under Chinese Policy

In Chinese building codes, it is required that residential buildings receive a minimum number of hours of natural, direct sunlight on a specified winter day, which represents the worst sunlight condition in a year. This requirement is a prerequisite for obtaining a building permit during the conceptual design of a residential project. Thus, officially sanctioned software is usually used to assess the sunlight performance of buildings. These software programs predict sunlight hours based on repeated shading calculations, which is time-consuming. This paper proposed a multilayer perceptron-based method, a one-stage prediction approach, which outputs a shading time interval caused by the inputted cuboid-form building. The sunlight hours of a site can be obtained by calculating the union of the sunlight time intervals (complement of shading time interval) of all the buildings. Three numerical experiments, i.e., horizontal level and slope analysis, and simulation-based optimization are carried out; the results show that the method reduces the computation time to 1/84~1/50 with 96.5%~98% accuracies. A residential neighborhood layout planning plug-in for Rhino 7/Grasshopper is also developed based on the proposed model. This paper indicates that deep learning techniques can be adopted to accelerate sunlight hour simulations at the conceptual design phase