Zhang et al. identify CEP55 as a novel regulator of cilia disassembly. Their findings establish a cilia disassembly role for CEP55 in vivo and in vitro, coupling defects in cilia disassembly to ciliopathy and further suggesting that proper cilia dynamics are critical for mammalian development.

Microtubule attachments to spindle poles in yeast are flexible, which allows the microtubules to pivot. Fong et al. directly measure the pivoting flexibility of the microtubule–pole interface and show that this flexibility is important for timely pole separation during mitosis.

Thaller et al. demonstrate that direct binding between phosphatidic acid (PA) and the ESCRT Chm7 is required for nuclear envelope surveillance; PA also accumulates at nuclear envelope herniations. Thus, tight control of PA metabolism is required for nuclear envelope homeostasis.

Kanfer et al. develop a pooled CRISPRi screening method to identify genes regulating intracellular protein localization, organelle morphology or other subcellular phenotypes. The method uses machine learning to identify genetically altered cells, a photoactivated fluorescent protein to label them, and FACS plus deep sequencing to identify the affected gene.

Yan et al. demonstrate high-throughput screening of pooled CRISPR libraries for phenotypes detectable by microscopy. Their approach uses photoactivation of cells displaying the phenotype of interest and FACS sorting of marked cells, followed by sequencing, and facilitates discovery of genes involved in cell biological processes.

The AP-5/SPG11/SPG15 complex is recruited onto late endosomes/lysosomes and contributes to lysosomal homeostasis and autophagic lysosome reformation. Hirst et al. show that recruitment is by coincidence detection, requiring both phosphatidylinositol 3-phosphate and Rag GTPases, thus uncovering a link between AP-5/SPG11/SPG15 and the mTORC1 pathway.

During T cell development, genomic rearrangement must create a T cell receptor capable of transmitting signals. Allam et al. show that passage through the β-selection checkpoint requires the assembly of a platform to support TCR signaling, similar to the mature T cell immunological synapse.