Anchors aweigh: using shRNA TO MAP p150Glued functionality in anchoring microtubules at the centrosome

Dynactin is a multisubunit protein complex required for proper functioning of the microtubule motor, cytoplasmic dynein. Dynactin serves as a processivity factor for the motor as well as a cargo adaptor, allowing dynein to function in a wide array of cellular processes. Additionally, dynactin serves as a microtubule anchor. The p150Glued subunit of dynactin is of particular importance to these processes, as it possesses dynactin’s microtubule binding sequences, termed the CAP-Gly and Basic domains. These domains have differential affinities for microtubules, with CAP-Gly binding to microtubules with a higher affinity than Basic. By testing a set of shRNA plasmids with antisense sequences to the untranslated region of p150Glued we have found effective knockdown of the protein in COS-7 cells; future researchers can then introduce plasmids for p150Glued lacking either the CAP-Gly or Basic domains, or both, potentially showing a differential effect on anchoring, dependent upon which domain is present.