In situ structure determination of conformationally flexible targets using nextPYP Single-particle cryoelectron tomography (SP-CET) is an imaging technique capable of determining the structure of proteins in their cellular environment at high-resolution. nextPYP is a web-based application designed to streamline the SP-CET structure determination process and facilitate the analysis of conformational variability. Here we explain how to use nextPYP-based methods…
MiLoPYP: self-supervised molecular pattern mining and particle localization in situ Cryo-electron tomography (CET) allows the routine visualization of cellular landscapes in three dimensions at nanometer-range resolutions. When combined with single-particle tomography (SPT), it is possible to obtain near-atomic resolution structures of frequently occurring macromolecules within their native environment. Two outstanding challenges associated with CET/SPT are the automatic identification and…
Redox-sensitive E2 Rad6 controls cellular response to oxidative stress via K63-linked ubiquitination of ribosomes In this study, we set out to investigate the key role of Rad6 in regulating cellular response to stress in budding yeast as part of the RTU. Rad6 is small (20 kDa), highly conserved, and a multifunctional E2 involved in DNA repair and in the…
Structural impact of K63 ubiquitin on yeast translocating ribosomes under oxidative stress K63 ubiquitination of ribosomes serves as a key regulator of protein production during cellular exposure to oxidative stress. Defining the structural and functional mechanisms of translation regulation would support the current understanding of critical reprogramming of eukaryotic gene expression. Our paper presents an examination of the structure…
