Analysis of Interactions between Regulator of G-Protein Signaling-14 and Microtubules

RGS14 is a Microtubule-Associated Protein

“…Notably, GST-RGS14-His 6 , purified as a soluble protein, appears to have a higher specific activity (GDI IC 50 = 164 nM) than the refolded protein (GDI IC 50 = 540 nM). 40 Thus, these data validate the purification of functionally active RGS14 for use in microtubule interaction and polymerization studies described hereafter.…”

“…We have described the isolation and refolding of the recombinant protein. 40 In this report, we purify GST-RGS14-His 6 as soluble protein from E. coli, using a three-column purification strategy (Fig. 3A).…”

Regulators of G-Protein-Signaling Proteins: Negative Modulators of G-Protein-Coupled Receptor Signaling

“…Further studies demonstrated that RGS14 interacts with and modulates the GTPSase activity of subunits of heterotrimeric proteins from G i/o families (Cho et al, 2000;Hepler et al, 2005;Traver et al, 2000). Actually, it is well known that RGS14 is implicated in a number of cellular processes, including, lymphocyte functions, centrosome formation and nuclear functions, and stress-induced signaling pathways (Cho and Kehrl, 2007;Cho et al, 2005Cho et al, , 2000Lin et al, 2011;Martin-McCaffrey et al, 2004a;Martin-McCaffrey et al, 2004b;MartinMcCaffrey et al, 2005b;Shu et al, 2007).…”

“…It regulates spindle formation, stabilizes, and is a component of mitotic aster. It has been shown that RGS14 enhances the GTPase activity of G ai2 , G ai3 , and G ai3 , promoting microtubule association in a GTP-dependent manner (Cho and Kehrl, 2007;Martin-McCaffrey et al, 2004b;Martin-McCaffrey et al, 2005b). Depletion of RGS14 from cell impaired aster formation (Martin-McCaffrey et al, 2005b) and results in cytofragmentation and failure to progress to the two-cell stage (MartinMcCaffrey et al, 2004a).…”

The Ras‐binding domain region of RGS14 regulates its functional interactions with heterotrimeric G proteins

“…RGS12 and RGS14 are among the largest RGS proteins, while the remaining D/R12 member, RGS10, is similar in size to the B/R4 subfamily of RGS proteins (Ross and Wilkie, 2000). Most studies on the physiological function of RGS14 have focused on its roles in the brain and in cell division (Lee et al, 2010; Martin-McCaffrey et al, 2004a; Martin-McCaffrey et al, 2004b; Martin-McCaffrey et al, 2005; Rodriguez-Munoz et al, 2007). For example, RGS14 is a mitotic spindle protein that associates with microtubules (Martin-McCaffrey et al, 2004a; Martin-McCaffrey et al, 2004b; Martin-McCaffrey et al, 2005).…”