SPECC1L binds the myosin phosphatase complex MYPT1/ PP1β and can regulate its distribution between microtubules and filamentous actin

The subcellular localization, activity , and substrate speci-ficity of the serine/threonine protein phosphatase 1 catalytic subunit (PP1cat) is mediated through its dynamic association with regulatory subunits in holoenzyme complexes. While some functional overlap is observed for the three human PP1cat isoforms, they also show distinct targeting based on relative preferences for specific regulatory subunits. A well-known example is the preferential association of MYPT1 with PP111 in the myosin phosphatase complex. In smooth muscle, MYPT1/PP111 counteracts the muscle contraction induced by phosphorylation of the light chains of myosin by the myosin light chain kinase. This phosphatase complex is also found in nonmuscle cells, where it is targeted to both myosin and nonmyosin substrates and contributes to regulation of the balance of cytoskeletal structure and motility during cell migration and division. Although it remains unclear how MYPT1/PP111 traffics between microtubule-and actin -associated substrates, our identification of the microtubule-and actin-binding protein SPECC1L in both the PP111 and MYPT1 interactomes suggests that it is the missing link. Our validation of their association using coimmunoprecipitation and proximity biotinylation assays, together with the strong overlap that we observed for the SPECC1L and MYPT1 inter-actomes, confirmed that they exist in a stable complex in the cell. We further showed that SPECC1L binds MYPT1 directly and that it can impact the balance of the distribution of the MYPT1/PP111 complex between the microtubule and filamen-tous actin networks.

Automated summary

The serine/threonine protein phosphatase 1 catalytic subunit (PP1cat) functions in subcellular localization, activity, and substrate specificity through its interaction with regulatory subunits. The PP1cat isoforms show distinct targeting based on their preferences for specific regulatory subunits. MYPT1/PP111, for example, preferentially associates with each other in the myosin phosphatase complex to counteract muscle contraction. This complex also plays a role in regulating cytoskeletal structure and motility in nonmuscle cells. The protein SPECC1L is identified as a potential link between microtubule-and actin-associated substrates for the MYPT1/PP111 complex.