Adaptor Autoregulation Promotes Coordinated Binding within Clathrin Coats

Abstract

Background: There are multiple interacting clathrin adaptors at the trans-Golgi network and endosomes in yeast. Results: Autoregulation of one adaptor, Gga2, alters the temporal delay between recruitment of Gga2 and a second adaptor, Ent5, to organelles. Conclusion:The interaction between Gga2 and Ent5 is regulated by an autoregulatory sequence. Significance: This autoregulatory mechanism may ensure accurate membrane traffic in vivo.

Energy metabolism regulates clathrin adaptors at thetrans-Golgi network and endosomes

“…, 2012). Furthermore, the Gga/Ent3 module promotes the recruitment of the Ent5/AP-1 module (Daboussi et al ., 2012; Hung et al ., 2012). Deletion of SNF1 did not dramatically alter adaptor localization in the presence of glucose or adaptor redistribution to the cytosol in the acute phase of glucose starvation (Figure 1, B and C).…”

Adaptor protein complex-1 (AP-1) is recruited by the HEATR5 protein Laa1 and its co-factor Laa2 in yeast

“…Alexa Fluor 647 goat anti-rabbit (RRID:AB_141663) antibody was from Life Technologies. Antibodies against Apl2, Ent5, and GST were described previously (44,45).…”

“…Cell lysates for GST-tagged protein interaction studies were generated by liquid nitrogen blending as previously described (44). Powder was thawed at a ratio of 0.15 g of lysate to 0.85 ml of HEKG5 with protease inhibitor mixture without EDTA (Promega).…”

“…The lysate was clarified by centrifugation at 13,000 rpm for 10 min, and the resulting supernatants were used for binding studies. Purification of GST and ␥-ear constructs were described previously (44). His-tagged Laa2 was purified using Talon resin according to the manufacturer's instructions (Clontech).…”

Direct binding of the Kex2p cytosolic tail to the VHS domain of yeast Gga2p facilitates TGN to prevacuolar compartment transport and is regulated by phosphorylation

“…The only other motif of at least 5 residues conserved between the Gga1p and Gga2p hinge domains, NLIDF (residues 351-355 in Gga2p), has recently been shown to contain overlapping GAE-domain and clathrin-binding motifs. This sequence governs a distinct autoinhibitory mechanism that regulates competing interactions of Gga2p with clathrin and the Ent5p adaptor (Hung et al, 2012). Thus there may be multiple internal mechanisms that regulate the multivalent binding activities of the yeast Gga proteins.…”