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Associate teacher rohan teasdale The endosomal system of mammalian cells is a very dynamic organelle, and the trafficking pathways associated with it are fundamental for many cellular processes.We are developing cellular and computational approaches to identify novel mammalian proteins linked to the endosomal system including the retromer protein complex that is a central regulator of early endosome protein trafficking.Retromer has been recently implicated in both alzheimer's and parkinson's neurological diseases and we are checking out the molecular mechanisms underlying these diseases.Macropinocytosis is a regulated form of endocytosis that mediates the non not bothered uptake of extracellular material.Numerous infectious pathogens exploit macropinocytosis to invade the host and we are currently investigation how it is modulated during pathogen infection. Endosomal dynamics and virus invasion The endo lysosomal system of mammalian cells offers highly dynamic endocytic membrane transport pathways that are fundamental for many cellular processes, incorporating receptor signalling, uptake of important nourishment and the ability to mount an effective lymphocyte response to foreign antigens.Important, defects in endosomal trafficking are related to many human diseases including various cancers, lysosomal storage health problems, neurodegenerative health-Related and hypercholesterolemia.Nearly, numerous infectious pathogens exploit specific endocytic pathways to attack the host.Characterisation of pathogen entry pathways is essential for appreciation infectious diseases but has also proven to be a powerful tool for gaining insight into normal cellular processes.As an example, to enhance survival, salmonella typhimurium subverts the host cell's normal macropinocytic machinery to gain entry into the non phagocytic epithelial cells of the digestive tract wall.Once inside cell, s.Typhimurium actively alters the phosphoinositide composition of the enveloping macropinosome to avert being delivered to the lysosome for destruction.For that reason, s.Typhimurium effectively manipulates the host cell's membrane trafficking pathways in order to make a replicative niche within a customised macropinosome.Knowledge of the cell biology of these clinically important intracellular pathogens and how they communicate with the host is incomplete.One of the most basic and intriguing facets of endosomal function is the mechanism of biomolecular transport, both within the endosomal system and back other cellular organelles.From your that the cell precisely control the spatial and temporal trafficking of these numerous individual proteins.A predominant strategy exploited by intracellular pathogens is to disrupt these transport pathways to ensure survival by directly targeting host proteins involved in membrane transport and agency of the endosome.My multi disciplinary research group is focused on focusing on how individual proteins are compartmentalized within the endo lysosomal system in mammalian cells, and defining the protein machinery regarding their organization and trafficking.We are currently looking the molecular details of these host pathways and how they are modulated in response to numerous infectious intracellular bacteria.We established high content screening(Hcs)Techniques, combined with high throughput rnai and automated imaging to enable the full concept of host pathogen interactions.Upon binding to the host cell spot, the pathogen utilises a spe******t apparatus called the type iii secretion(T3ss)System to deliver a host of bacterial effector proteins straight to the host cell's cytoplasm.These virulence proteins directly manipulate the infected cell's cytoskeletal construction, membrane layer trafficking, signal transduction and cytokine gene expression to promote bacterial duplication and survival within the host's cells.To establish the network of host proteins that communicate with salmonella effectors secreted into the host cells we have cloned all salmonella effector proteins defined to date.We will methodically determine the host proteins they interact with using co immunoprecipitation and mass spectrometry/proteomics.The advantages of these identified host proteins, based on myself and others, for salmonella duplication or invasion will be examined using rnai knockdown of the host proteins. Defining the role host cell membrane trafficking pathways play in chlamydial introduction formation. Chlamydia are obligate intracellular pathogens regarding a range of human and animal diseases.To live within the host cell, the pathogen pirates the host's membrane trafficking pathways to engineer an intracellular niche called an introduction.Besides providing a permissive environment, this plan also shields the pathogen from the host's immune system.Using established molecular tools in conjunction with sophisticated live cell imaging technology we will examine the membrane trafficking pathways hijacked by the pathogen and specifically key regulators of these pathways. Tailor-Made endosomal pathways are emerging as distinct critical biological systems present within restricted cell types.Such as the high capacity macropinocytosis, the virus induced phagocytosis and clathrin independent endocytosis via caveolae.Rab gtpases)But clearly each have additional unique regulatory intricacies.Macropinocytosis represents a regulated form of endocytosis that can be induced by growth factors and has developed into recent focus of my group. Characterisation of the px domain necessary family in macropinocytosis. Apart from their seriousness as signalling molecules, phosphoinositides are required for spatio temporal recruitment of protein effectors, because phox homology(Px)Domain individual.The mammalian genome encodes forty nine proteins that possess a px domain with regard to membrane attachment to organelles of the secretory and endocytic system via binding of phosphoinositide lipids.Nearly all px domain proteins bind the phosphoinositide ptdins3p and are recruited to early endosomal membranes.The px domain proteins most of which are grouped as sorting nexins(Snxs)Are emerging as vital endosome associated proteins, doing the job as trafficking cargo adaptors and membrane associated scaffolds regulating cell signalling.Many px proteins are linked to diseases and genetic disorders, defined within pathogen invasion, cancer and nerve disease.Using shrna controlling, cutting edge live cell imaging and biochemistry, we characterized phosphoinositide binding protein network of snxs during macropinocytosis and focused on proteins required for invasion and replication of s.Typhimurium a model intracellular pathogen which subverts phosphoinositide metabolic process, actin polymerization machines and membrane trafficking in the host cells via secreted effectors. Required peptids trafficking in disease The regulated movement of membrane receptors and ligands between the cell surface and intracellular pockets is vital to many cellular operations, including communicating between cells and their environment.I have a certain research program characterising the mammalian endosomal protein complex, termed the retromer area.A short while ago, the mammalian retromer protein complex can have a central role in the sorting of protein cargo within the endosome.Mammalian retromer is based on three proteins, vps29, vps35 with vps26, which form a high appreciation protein complex.Retromer is crucial for endosomal trafficking, get hold of for the retrograde transport of receptors from the endosome to the golgi.Retromer is emerging as something of the endosomal membrane trafficking and cargo sorting machineries. Retromer's role in adipocyte difference and glut 4 trafficking. At adipocytes, the blood sugar transporter, glut4 undergoes a regulated endosomal trafficking event in step to extracellular insulin.This is done by the generation of a unique endosomal compartment, called gsvs, in which glut 4 is controlled intracellular in resting cells.Until now, the role of retromer in the retention of gsvs within classified adipocytes is uncharacterized.We united states that the distinct vps26 subunits(A also known as b)Control the cargo binding properties of retromer and identified that the vps26b subunit of retromer is regulated upon difference of 3t3l1 cells into adipocytes.We will determine the need for retromer in gsvs function and will examine the differential roles of ubiquitous vps26a and adipocyte specific vps26b retromer in this process.This will represent the first bodily function for vps26b retromer to be described. Retromer's role in nerve disease. Alzheimer'sDiseaseDisease(Text marketing)And parkinson's sickness(Pd)Are progressive neurologicalDisorders that are reasons for causes ofDementia.Retromer was implicated in ad after the watching with interest that it isDown regulated in the brains of ad patients.TheDirect role of vps35 in quelling ad was confirmedUsing a hemizygousDeletion of vps35 in an ad mouse model.In addition, two recent surveys show several point mutations of vps35 cause late onset pd.Reports implicating vps35 in pd teach you a single point mutation of highly conserved amino acid 620(Asp620asn)In several families leads to an autosomalDominant mode of pd monetary gift.ToDate little isUnderstood about the mode of action of retromer in these nerveDiseases.In venture withDr.U.Coulson(QBI,UQ)And simplyDr.D.Collins(IMB,UQ) We have commenced studies to pick which of the known cellular or biochemical properties of Retromer contribute to the molecular mechanismsUnderlying theseDisease states.Using a knock out/knock in approach we will examine a number of structure based Retromer mutantsDeficient in individual biochemical properties and the PdDisease causing mutants in cell models and animal models. Magazine article:The globally displayed m1t1 clone of group a streptococcus evades autophagy for intracellular replication Barnett, timothy h, liebl, mark, seymour, lisa e, gillen, christine d, lim, jin yan, larock, captain christopher n, davies, indicate r, schulz, benjamin d, nizet, winner, teasdale, rohan chemical.And runner, note j. (2013)The globally displayed m1t1 clone of group a streptococcus evades autophagy for intracellular replication. Cell host and microorganisms, 14 6:675 682. Log article:The vps35 d620n mutation in connection with parkinson disease disrupts the cargo sorting function of retromer Log article:Architectural basis for endosomal trafficking of diverse transmembrane cargos by px ferm proteins Ghai, rajesh, bugarcic, andrea, liu, huadong, norwood, suzanne l, skeldal, sune, coulson, electronic j, li, shawn avoid cheng, teasdale, rohan ve hag.And as well, collins, brett d. (2013)Architectural basis for endosomal trafficking of diverse transmembrane cargos by px ferm proteins.

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