Sec14, the major yeast phosphatidylinositol (PtdIns)/phosphatidylcholine (PtdCho) transfer protein, regulates essential interfaces between lipid metabolism and membrane trafficking from the trans-Golgi network (TGN). How Sec14 does so remains unclear. We report that Sec14 binds PtdIns and PtdCho at distinct (but overlapping) sites, and both PtdIns- and PtdCho-binding activities are essential Sec14 activities. We further show both activities must reside within the same molecule to reconstitute a functional Sec14 and for effective Sec14-mediated regulation of phosphoinositide homeostasis in vivo. This regulation is uncoupled from PtdIns-transfer activity and argues for an interfacial presentation mode for Sec14-mediated potentiation of PtdIns kinases. Such a regulatory role for Sec14 is a primary counter to action of the Kes1 sterol-binding protein that antagonizes PtdIns 4-OH kinase activity in vivo. Collectively, these findings outline functional mechanisms for the Sec14 superfamily and reveal additional layers of complexity for regulating phosphoinositide homeostasis in eukaryotes.

Practical application of hybrid Pb-based halide perovskites needs not only to fabricate high-quality film samples on a large scale but also to properly overcome the issues of Pb toxicity and materials instability. Finding new, stable, Pb-free perovskites currently attracts significant research interest. Among various strategies, hetero-substitution of Pb to form quaternary halide double perovskites represents a promising direction to keep the high structural (and likely electronic) dimensionality nature of perovskite lattice and meanwhile offers rich chemical compositions a degree of freedom for discovering new perovskite materials. We herein present a perspective that concisely reviews the progress of rational design of Pb-free halide double perovskites by both theoretical and experimental efforts as well as their current and potential applications in various optoelectronic categories. We also envision the future research directions to realize new materials by exploring broader chemical composition space and to better utilize existing materials by considering optoelectronic properties modulation.

Function of the heart begins long before its formation is complete. Analyses in mouse and zebrafish have shown that myocardial function is not required for early steps of organogenesis, such as formation of the heart tube or chamber specification. However, whether myocardial function is required for later steps of cardiac development, such as endocardial cushion (EC) formation, has not been established. Recent technical advances and approaches have provided novel inroads toward the study of organogenesis, allowing us to examine the effects of both genetic and pharmacological perturbations of myocardial function on EC formation in zebrafish. To address whether myocardial function is required for EC formation, we examined silent heart (sih−/−) embryos, which lack a heartbeat due to mutation of cardiac troponin T (tnnt2), and observed that atrioventricular (AV) ECs do not form. Likewise, we determined that cushion formation is blocked in cardiofunk (cfk−/−) embryos, which exhibit cardiac dilation and no early blood flow. In order to further analyze the heart defects in cfk−/− embryos, we positionally cloned cfk and show that it encodes a novel sarcomeric actin expressed in the embryonic myocardium. The Cfks11 variant exhibits a change in a universally conserved residue (R177H). We show that in yeast this mutation negatively affects actin polymerization. Because the lack of cushion formation in sih- and cfk-mutant embryos could be due to reduced myocardial function and/or lack of blood flow, we approached this question pharmacologically and provide evidence that reduction in myocardial function is primarily responsible for the defect in cushion development. Our data demonstrate that early myocardial function is required for later steps of organogenesis and suggest that myocardial function, not endothelial shear stress, is the major epigenetic factor controlling late heart development. Based on these observations, we postulate that defects in cardiac morphogenesis may be secondary to mutations affecting early myocardial function, and that, in humans, mutations affecting embryonic myocardial function may be responsible for structural congenital heart disease.

Vps4p (End13p) is an AAA-family ATPase that functions in membrane transport through endosomes, sorting of soluble vacuolar proteins to the vacuole, and multivesicular body (MVB) sorting of membrane proteins to the vacuole lumen. In a yeast two-hybrid screen with Vps4p as bait we isolated VPS20 (YMR077c) and the novel open reading frame YLR181c, for which the name VTA1 has recently been assigned (Saccharomyces Genome Database). Vps4p directly binds Vps20p and Vta1p in vitro and binding is not dependent on ATP - conversely, Vps4p binding to Vps20p is partially sensitive to ATP hydrolysis. Both ATP binding [Vps4p-(K179A)] and ATP hydrolysis [Vps4p-(E233Q)] mutant proteins exhibit enhanced binding to Vps20p and Vta1p in vitro. The Vps4p-Vps20p interaction involves the coiled-coil domain of each protein, whereas the Vps4p-Vta1p interaction involves the (non-coiled-coil) C-terminus of each protein. Deletion of either VPS20 (vps20Δ) or VTA1 (vta1Δ) leads to similar class E Vps- phenotypes resembling those of vps4Δ, including carboxypeptidase Y (CPY) secretion, a block in ubiquitin-dependent MVB sorting, and a delay in both post-internalisation endocytic transport and biosynthetic transport to the vacuole. The vacuole resident membrane protein Sna3p (whose MVB sorting is ubiquitin-independent) does not appear to exit the class E compartment or reach the vacuole in cells lacking Vps20p, Vta1p or Vps4p, in contrast to other proteins whose delivery to the vacuole is only delayed. We propose that Vps20p and Vta1p regulate Vps4p function in vivo.

Incorporation of two-dimensional (2D) materials in electronic devices inevitably involves contact with metals, and the nature of this contact (Ohmic and/or Schottky) can dramatically affect the electronic properties of the assembly. Controlling these properties to reliably form low-resistance Ohmic contact remains a great challenge due to the strong Fermi level pinning (FLP) effect at the interface. Herein, we employ density functional theory calculations to show that van der Waals stacking can significantly modulate Schottky barrier heights in the contact formed between multilayer InSe and 2D metals by suppressing the FLP effect. Importantly, the increase of InSe layer number induces a transition from Schottky to Ohmic contact, which is attributed to the decrease of the conduction band minimum and rise of the valence band maximum of InSe. Based on the computed tunneling and Schottky barriers, Cd 3 C 2 is the most compatible electrode for 2D InSe among the materials studied. This work illustrates a straightforward method for developing more effective InSe-based 2D electronic nanodevices.

The Sec14-like phosphatidylinositol transfer protein Sfh3 associates with bulk LDs in vegetative cells but targets to a neutral lipid hydrolase-rich LD pool during sporulation. Sfh3 inhibits LD utilization by a PtdIns-4-phosphate–dependent mechanism, and this inhibition prevents prospore membrane biogenesis in sporulating cells.

Ubiquitinated integral membrane proteins are delivered to the interior of the lysosome/vacuole for degradation. This process relies on specific ubiquitination of potential cargo and recognition of that Ub-cargo by sorting receptors at multiple compartments. We show that the endosomal Hse1-Vps27 sorting receptor binds to ubiquitin peptidases and the ubiquitin ligase Rsp5. Hse1 is linked to Rsp5 directly via a PY element within its C-terminus and through a novel protein Hua1, which recruits a complex of Rsp5, Rup1, and Ubp2. The SH3 domain of Hse1 also binds to the deubiquitinating protein Ubp7. Functional analysis shows that when both modes of Rsp5 association with Hse1 are altered, sorting of cargo that requires efficient ubiquitination for entry into the MVB is blocked, whereas sorting of cargo containing an in-frame addition of ubiquitin is normal. Further deletion of Ubp7 restores sorting of cargo when the Rsp5:Hse1 interaction is compromised suggesting that both ubiquitin ligases and peptidases associate with the Hse1-Vps27 sorting complex to control the ubiquitination status and sorting efficiency of cargo proteins. Additionally, we find that disruption of UBP2 and RUP1 inhibits MVB sorting of some cargos suggesting that Rsp5 requires association with Ubp2 to properly ubiquitinate cargo for efficient MVB sorting. INTRODUCTIONIntegral membrane proteins are sorted into intralumenal vesicles of endosomes before delivery and degradation in lysosomes (Gruenberg and Stenmark, 2004). One of the sorting signals for sending proteins into the endosomal lumen is ubiquitin (Ub; Katzmann et al., 2002;Hicke and Dunn, 2003). Many studies have demonstrated that monoubiquitination of cargo is sufficient for efficient sorting into the interior of multivesiculated endosomes/bodies (MVB). Ubiquitination works as an MVB-sorting signal for a variety of proteins, including those internalized from the cell surface as well as those delivered to endosomes from the Golgi apparatus via the biosynthetic pathway. Ub also serves as a sorting signal at other post-Golgi compartments with the cumulative effect of sending cargo toward the lysosome. Ub can serve as a signal for internalization, thus hastening the delivery of cell surface proteins to endosomes, and Ub addition can also sort membrane proteins at the trans-Golgi network (TGN), diverting them to endosomes and preventing their delivery to the cell surface (Staub and Rotin, 2006).Ubiquitin works as a sorting signal by binding Ub-sorting receptors that recognize ubiquitinated cargo (Ub-cargo) and incorporate it into various transport intermediates (Staub and Rotin, 2006). At the MVB, proteins such as the Hrs-STAM (hepatocyte receptor substrate-signal transducing adaptor molecule) complex and the yeast Hse1-Vps27 complex bind Ub-cargo using multiple UIMs (ubiquitin interaction motifs) and help sort cargo into lumenal vesicles. At the TGN, the monomeric clathrin-associated GGA (Golgi-localized, gamma-ear containing, Arf-binding) proteins bind Ub via a GAT (GGA and TOM1) domain to faci...

Ubiquitin (Ub) is a sorting signal that targets integral membrane proteins to the interior of the vacuole/lysosome by directing them into lumenal vesicles of multivesicular bodies (MVBs). The Vps27-Hse1 complex, which is homologous to the Hrs-STAM complex in mammalian cells, serves as a Ubsorting receptor at the surface of early endosomes. We have found that Hse1 interacts with Doa1/Ufd3. Doa1 is known to interact with Cdc48/p97 and Ub and is required for maintaining Ub levels. We find that the Hse1 Src homology 3 domain binds directly to the central PFU domain of Doa1. Mutations in Doa1 that block Hse1 binding but not Ub binding do not alter Ub levels but do result in the missorting of the MVB cargo GFP-Cps1. Loss of Doa1 also causes a synthetic growth defect when combined with loss of Vps27. Unlike the loss of Doa1 alone, the doa1⌬ vps27⌬ double mutant phenotype is not suppressed by Ub overexpression, demonstrating that the effect is not due to indirect consequence of lowered Ub levels. Loss of Doa1 results in a defect in the accumulation of GFP-Ub within yeast vacuoles, implying that there is a reduction in the flux of ubiquitinated membrane proteins through the MVB pathway. This defect was also reflected by an inability to properly sort Vph1-GFP-Ub, a modified subunit of the multiprotein vacuolar ATPase complex, which carries an inframe fusion of Ub as an MVB sorting signal. These results reveal novel roles for Doa1 in helping to process ubiquitinated membrane proteins for sorting into MVBs.