DYNLT (Tctex‐1) forms a tripartite complex with dynein intermediate chain and RagA, hence linking this small GTP ase to the dynein motor

• Published in: The FEBS journal 2015-10, Vol.282 (20), p.3945-3958
• Main Authors: Merino‐Gracia, Javier, García‐Mayoral, María Flor, Rapali, Peter, Valero, Ruth Ana, Bruix, Marta, Rodríguez‐Crespo, Ignacio
• Format: Article
• Language: English
• ISSN: 1742-464X; 1742-4658
• DOI: 10.1111/febs.13388

It has been suggested that DYNLT , a dynein light chain known to bind to various cellular and viral proteins, can function as a microtubule–cargo adaptor. Recent data showed that DYNLT links the small GTP ase Rab3D to microtubules and, for this to occur, the DYNLT homodimer needs to display a binding site for dynein intermediate chain together with a binding site for the small GTP ase. We have analysed in detail how RagA, another small GTP ase, associates to DYNLT . After narrowing down the binding site of RagA to DYNLT we could identify that a β strand, part of the RagA G3 box involved in nucleotide binding, mediates this association. Interestingly, we show that both microtubule‐associated DYNLT and cytoplasmic DYNLT are equally able to bind to the small GTP ases Rab3D and RagA. Using NMR spectroscopy, we analysed the binding of dynein intermediate chain and RagA to mammalian DYNLT . Our experiments identify residues of DYNLT affected by dynein intermediate chain binding and residues affected by RagA binding, hence distinguishing the docking site for each of them. In summary, our results shed light on the mechanisms adopted by DYNLT when binding to protein cargoes that become transported alongside microtubules bound to the dynein motor.