In fungal hyphal cells, intracellular membrane trafficking is constrained by the relatively long distances and mode of growth, exclusively apical extension. Endocytosis plays a key role in tip which involves coupling secretory delivery to region with subapical compensatory endocytosis. However, identity, dynamics function filamentous endosomal compartments remain largely unexplored. Aspergillus nidulans RabA(Rab5) localizes population endosomes that show range bidirectional movement on...

The use of Drosophila melanogaster as a model organism has been pivotal to understanding the developmental processes metazoans. However, flies for studying subcellular organization is hampered by paucity reliable reagents label specific organelles. Here, we describe generation mouse monoclonal antibodies against set markers secretory and endocytic pathways, along with goat polyclonal two Golgi proteins. We show that are highly sufficiently sensitive detect endogenous proteins in crude...

Originally identified in yeast, transport protein particle (TRAPP) complexes are Rab GTPase exchange factors that share a core set of subunits. TRAPPs were initially found to act on Ypt1, the yeast orthologue Rab1, but recent studies have TRAPPII can also activate Rab11 orthologues Ypt31/32. Mammals two TRAPP complexes, their role is less clear, and they contain subunits not essential for cell growth. To investigate function metazoans, we show Drosophila melanogaster similar those mammals...

Aspergillus nidulans early endosomes display characteristic long-distance bidirectional motility. Simultaneous dual-channel acquisition showed that the two Rab5 paralogues RabB and RabA colocalize in these also larger, immotile mature endosomes. However, RabB-GTP is sole recruiter to of Vps34 PI3K (phosphatidylinositol-3-kinase) phosphatidylinositol-3-phosphate [PI(3)P] effector AnVps19 rabB Delta, leading thermosensitivity prevents multivesicular body sorting endocytic cargo. Thus, mediator...

The fungal pal/RIM signalling pathway, which regulates gene expression in response to environmental pH involves, addition dedicated proteins, several components of ESCRT complexes, suggested that proteins assemble on endosomal platforms. In Aspergillus nidulans, Pal include the plasma membrane receptor PalH and its coupled arrestin, PalF, becomes ubiquitylated alkaline conditions, three potentially ESCRT-III associates, including Vps32 interactors PalA PalC Vps24 interactor calpain-like...

The genetically tractable filamentous ascomycete fungus Aspergillus nidulans has been successfully exploited to gain major insight into the eukaryotic cell cycle. More recently, its amenability in vivo multidimensional microscopy fueled a potentially gilded second age of A. biology studies. This review specifically deals with studies on intracellular membrane traffic nidulans. cellular logistics are subordinated needs imposed by polarized mode growth multinucleated hyphal tip cells, whereas...

We exploit the ease with which highly motile early endosomes are distinguished from static late in order to study Aspergillus nidulans endosomal traffic. RabS(Rab7) mediates homotypic fusion of endosomes/vacuoles a fusion- and vacuole protein sorting/Vps41-dependent manner. Progression across endocytic pathway involves maturation because end products absence minivacuoles that competent multivesicular body sorting cargo degradation but retain features, such as ability undergo long-distance...

The TRAPP complexes are nucleotide exchange factors that play essential roles in membrane traffic and autophagy. TRAPPII activates Rab11, TRAPPIII Rab1, with the two sharing a core of small subunits affect but being distinguished by specific large for activity vivo. Crystal structures have revealed mechanism Rab activation, how assemble to form is unknown. We report cryo-EM structure entire Drosophila complex. TRAPPIII-specific TRAPPC8 TRAPPC11 hold catalytic like pair tongs, TRAPPC12...

PalC, distantly related to Saccharomyces cerevisiae peripheral endosomal sorting complexes required for transport III (ESCRT‐III) component Bro1p and one of six Aspergillus nidulans pH signalling proteins, contains a Bro1 domain. Green fluorescent protein (GFP)‐tagged PalC is recruited plasma membrane‐associated punctate structures upon alkalinization, when active. recruitment these dependent on the seven transmembrane domain (7‐TMD) receptor likely sensor PalH. two‐hybrid interactor...

The seven-transmembrane receptor PalH and its coupled, positive-acting arrestin-like protein PalF are key components of a molecular sensor that in Aspergillus nidulans other ascomycete fungi mediates activation an intracellular signaling cascade by alkaline ambient pH. is ubiquitinated pH- PalH-dependent manner. We show here assists the plasma membrane localization overexpression slightly hypersensitizes pathway to pH but does not bypass need for signal signaling. In contrast, covalent...

Activation of the Aspergillus nidulans transcription factor PacC, which mediates ambient pH regulation gene expression and is recruited to ESCRT-III by Vps32-interacting scaffold PalA, involves its pH-dependent C-terminal proteolysis. This reaction almost certainly catalyzed PalB calpain-like protease. Here we show that associates with membranes interacts specifically directly Vps24. The N-terminal MIT domain Vps24 MIM motif are necessary sufficient for this interaction. PalB(DeltaMIT), a...

The Aspergillus pal pathway hijacks ESCRT proteins into ambient pH signalling complexes. We show that components of ESCRT-0, ESCRT-I, ESCRT-II and ESCRT-III are nearly essential for growth, precluding assessment null mutants or trafficking. This severely debilitating effect is rescued by loss-of-function mutations in two cation tolerance genes, one which, sltA, encodes a transcription factor whose inactivation promotes hypervacuolation. Exploiting conditional expression sltA allele, we...

The GTPase Arf1 is the major regulator of vesicle traffic at both cis- and trans-Golgi. activated cis-Golgi by guanine nucleotide exchange factor (GEF) GBF1 trans-Golgi related GEF BIG1 or its paralog, BIG2. trans-Golgi-specific targeting BIG2 depends on Arf-like Arl1. We find that Arl1 binds to dimerization cyclophilin binding (DCB) domain in report a crystal structure human bound this domain. Residues DCB bind are required for locate Golgi vivo. domain-binding residues have distinct...

ABSTRACT Aspergillus nidulans (Pal) ambient pH signaling takes place in cortical structures containing components of the ESCRT pathway, which are hijacked by alkaline pH-activated, ubiquitin-modified version arrestin-like protein PalF and taken to plasma membrane. There, ESCRTs scaffold assembly dedicated Pal proteins acting downstream. The molecular details this results two-step proteolytic processing transcription factor PacC, have received considerable attention due key role that it plays...

Gallicin, a new germacranolide, was isolated from Artemisia maritima gallica Willd and assigned structure (3a) on chemical spectroscopic evidence. Its absolute configuration determined by the fact that it can be converted to dihydrosantamarin (5) biogenetic cyclization process.

Three new sesquiterpene lactones, linichlorins A, B, and C were isolated from the aerial parts of Centaurealinifolia Vahl their structures determined by physical chemical methods. The lactones chlorohyssopifolins C, D, E, aguerin vahlenin also same plant.

The pal/RIM ambient pH signalling pathway is crucial for the ability of pathogenic fungi to infect hosts. Aspergillus nidulans 7-TMD receptor PalH senses alkaline pH, subsequently facilitating ubiquitination arrestin PalF. Ubiquitinated PalF triggers downstream events. mechanism(s) by which transduces signal poorly understood. We show that phosphorylated in a dependent manner, resembling mammalian GPCRs, although phosphorylation, contrast dependent. A genetic screen revealed an...

Uso1/p115 and RAB1 tether ER-derived vesicles to the Golgi. contains a globular-head-domain (GHD), coiled-coil (CC) mediating dimerization/tethering, C-terminal region (CTR) interacting with golgins. is recruited by RAB1. Genetic studies placed Uso1 paradoxically acting upstream of, or in conjunction (Sapperstein et al., 1996). We selected two missense mutations uso1 resulting E6K G540S GHD that rescued lethality of rab1 -deficient Aspergillus nidulans . The are phenotypically additive,...

A synthetic strategy for the preparation of trisusbstituted cyclic ethers is presented, in which stereochemistry at carbon atoms adjacent to oxygen ether was controlled by means a hetero Diels−Alder reaction between monoactivated diene and chiral aldehyde. The adducts were transformed into linear ethers, then used cis trans different sizes. cis-oxepane, cis-oxonane, cis-oxocane trans-oxocane prepared as examples scope strategy.

In the apex-directed RAB11 exocytic pathway of Aspergillus nidulans, kinesin-1/KinA conveys secretory vesicles (SVs) to hyphal tip, where they are transferred type V myosin MyoE. MyoE concentrates SVs at an apical store located underneath PM resembling presynaptic active zone. A rod-shaped effector, UDS1, and intrinsically disordered coiled-coil HMSV associate with in a stable HUM (HMSV-UDS1-MyoE) complex recruited by through interaction network involving components, globular tail domain...

The oxidation of olefins and ketones to oxiranes esters, respectively, is carried out with the UHP (urea-hydrogen peroxide complex)maleic anhydride system in a mild very simple procedure.The epoxidation peroxyacids (MCPBA, PBA, 3,5-DNPBA, PAA, TIT 'Aq MPPA monoperoxymaleic acid) impoaant reaction organic synthesis.1Recently, Heaney et a1.2 have shown that magnesium monoperoxypthalate (MMPP), as well complex urea-hydrogen (UHP), together acetic trifluoroacetic anhydrides presence buffer...