TY - JOUR T1 - Lysate-based pipeline to characterize microtubule-associated proteins uncovers unique microtubule behaviours. JF - Nat Cell Biol Y1 - 2022 A1 - Jijumon, A S A1 - Bodakuntla, Satish A1 - Genova, Mariya A1 - Bangera, Mamata A1 - Sackett, Violet A1 - Besse, Laetitia A1 - Maksut, Fatlinda A1 - Henriot, Veronique A1 - Magiera, Maria M A1 - Sirajuddin, Minhajuddin A1 - Janke, Carsten AB -

The microtubule cytoskeleton forms complex macromolecular assemblies with a range of microtubule-associated proteins (MAPs) that have fundamental roles in cell architecture, division and motility. Determining how an individual MAP modulates microtubule behaviour is an important step in understanding the physiological roles of various microtubule assemblies. To characterize how MAPs control microtubule properties and functions, we developed an approach allowing for medium-throughput analyses of MAPs in cell-free conditions using lysates of mammalian cells. Our pipeline allows for quantitative as well as ultrastructural analyses of microtubule-MAP assemblies. Analysing 45 bona fide and potential mammalian MAPs, we uncovered previously unknown activities that lead to distinct and unique microtubule behaviours such as microtubule coiling or hook formation, or liquid-liquid phase separation along the microtubule lattice that initiates microtubule branching. We have thus established a powerful tool for a thorough characterization of a wide range of MAPs and MAP variants, thus opening avenues for the determination of mechanisms underlying their physiological roles and pathological implications.

VL - 24 IS - 2 ER -