The endoplasmic reticulum connects to the nucleus by constricted junctions that mature after mitosis

Abstract Junctions between the endoplasmic reticulum (ER) and the outer membrane of the nuclear envelope (NE) physically connect both organelles. These ER–NE junctions are essential for supplying the NE with lipids and proteins synthesized in the ER. However, little is known about the structure of these ER–NE junctions. Here, we systematically study the ultrastructure of ER–NE junctions in cryo-fixed mammalian cells staged in anaphase, telophase, and interphase by correlating live cell imaging with three-dimensional electron microscopy. Our results show that ER–NE junctions in interphase cells have a pronounced hourglass shape with a constricted neck of 7–20 nm width. This morphology is significantly distinct from that of junctions within the ER network, and their morphology emerges as early as telophase. The highly constricted ER–NE junctions are seen in several mammalian cell types, but not in budding yeast. We speculate that the unique and highly constricted ER–NE junctions are regulated via novel mechanisms that contribute to ER-to-NE lipid and protein traffic in higher eukaryotes.