Auxilin facilitates membrane traffic in the early secretory pathway

Ding, Jingzhen; Segarra, Verónica A.; Chen, Shuliang; Cai, Huaqing; Lemmon, Sandra K.; Ferro‐Novick, Susan

doi:10.1091/mbc.e15-09-0631

Cited by 16 publications

(17 citation statements)

References 49 publications

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“…First, similar to other cargo receptors it cycles between the ER and Golgi and is affected by Sec24 cargo‐binding site mutants. In support of our findings, Exp1 was found to be enriched in COPI/COPII vesicles in 2 independent studies and we found that it is packaged into COPII vesicles in vitro. Second, Exp1 physically binds Pma1 as would be expected for a cargo receptor.…”

Section: Discussionsupporting

confidence: 90%

Two novel effectors of trafficking and maturation of the yeast plasma membrane H⁺‐ATPase

Geva

Crissman

Arakel

et al. 2017

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The endoplasmic reticulum (ER) is the entry site of proteins into the endomembrane system.Proteins exit the ER via coat protein II (COPII) vesicles in a selective manner, mediated either by direct interaction with the COPII coat or aided by cargo receptors. Despite the fundamental role of such receptors in protein sorting, only a few have been identified. To further define the machinery that packages secretory cargo and targets proteins from the ER to Golgi membranes, we used multiple systematic approaches, which revealed 2 uncharacterized proteins that mediate the trafficking and maturation of Pma1, the essential yeast plasma membrane proton ATPase. Ydl121c (Exp1) is an ER protein that binds Pma1, is packaged into COPII vesicles, and whose deletion causes ER retention of Pma1. Ykl077w (Psg1) physically interacts with Exp1 and can be found in the Golgi and coat protein I (COPI) vesicles but does not directly bind Pma1. Loss of Psg1 causes enhanced degradation of Pma1 in the vacuole. Our findings suggest that Exp1 is a Pma1 cargo receptor and that Psg1 aids Pma1 maturation in the Golgi or affects its retrieval. More generally our work shows the utility of high content screens in the identification of novel trafficking components.The endoplasmic reticulum (ER) is the entry site for proteins into the endomembrane system. Once appropriately folded, non-ER resident proteins are routed to the Golgi apparatus for further maturation and distribution.1 Proteins exit the ER via coat protein II (COPII) vesicles.Although some proteins are captured by stochastic sampling of the ER lumen and membrane in a process called bulk flow, 2,3 more efficient and regulated sorting relies on selective uptake into vesicles.Additional sorting specificity occurs by retrieval of ER residents or immature proteins from the Golgi apparatus to the ER through coat protein I (COPI) vesicles. 4,5 The general mechanisms of vesicle formation are well characterized, yet the molecular basis that governs specific recognition between the coats, and the diverse range of proteins that must be packaged into vesicles, is yet to be fully described. To perturb traffic we chose to use mutations in the cargo binding sites of Sec24 as it was already shown that a mutation in the Sec24-A binding site perturbs ER-Golgi vesicular traffic. 26 Moreover, the 2 -best-characterized cargo of the A-and C-sites are SNARE proteins that act in anterograde (Sed5) and retrograde (Sec22) traffic, suggesting that bidirectional traffic between the ER and Golgi would be perturbed in these mutants. In contrast, many cargo that engage the Bsite are proteins that move forward in the secretory pathway. Thus we focused on A-and C-site mutations that should more specifically perturb ER-Golgi vesicular traffic. 6,7 We used automated mating techniques to integrate these Sec24 binding site mutants into the secretome-GFP library 27 and imaged the resulting strains using a high content setup ( Figure 1A).

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Section: Discussionsupporting

confidence: 90%

Two novel effectors of trafficking and maturation of the yeast plasma membrane H⁺‐ATPase

Geva

Crissman

Arakel

et al. 2017

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“…We were unable to express full length LdSec24 and LdSec31 in E. coli as observed previously (54). Therefore, LdSec31…”

Section: Generation and Purification Of Different Mutants Of Copii Prmentioning

confidence: 69%

GTPase Sar1 regulates the trafficking and secretion of the virulence factor gp63 in Leishmania

Parashar

Mukhopadhyay

2017

Journal of Biological Chemistry

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“…This is in contrast to loss-of-function alleles of other endocytic components, which exhibit various defects in growth, development, and/or reduced fertility (Gadeyne et al, 2014;Kang et al, 2001Kang et al, , 2003Collings et al, 2008;Kim et al, 2013;Kitakura et al, 2011). This observation also differs from that of auxilin loss-of-function mutants in yeast, whose growth is reduced (Ding et al, 2015), and auxilin/GAK lossof-function mutants in animals such as mouse, worm, or zebra fish, where various aspects of development are affected (Yim et al, 2010;Greener et al, 2001;Bai et al, 2010;Lee et al, 2008).…”

Section: Discussionmentioning

confidence: 88%

A Functional Study of AUXILIN-LIKE1 and 2, Two Putative Clathrin Uncoating Factors in Arabidopsis

et al. 2018

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Clathrin-mediated endocytosis (CME) is a cellular trafficking process in which cargoes and lipids are internalized from the plasma membrane into vesicles coated with clathrin and adaptor proteins. CME is essential for many developmental and physiological processes in plants, but its underlying mechanism is not well characterized compared with that in yeast and animal systems. Here, we searched for new factors involved in CME in by performing tandem affinity purification of proteins that interact with clathrin light chain, a principal component of the clathrin coat. Among the confirmed interactors, we found two putative homologs of the clathrin-coat uncoating factor auxilin previously described in non-plant systems. Overexpression of AUXILIN-LIKE1 and AUXILIN-LIKE2 in Arabidopsis caused an arrest of seedling growth and development. This was concomitant with inhibited endocytosis due to blocking of clathrin recruitment after the initial step of adaptor protein binding to the plasma membrane. By contrast, loss-of-function lines did not present endocytosis-related developmental or cellular phenotypes under normal growth conditions. This work contributes to the ongoing characterization of the endocytotic machinery in plants and provides a robust tool for conditionally and specifically interfering with CME in Arabidopsis.

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Auxilin facilitates membrane traffic in the early secretory pathway

Cited by 16 publications

References 49 publications

Two novel effectors of trafficking and maturation of the yeast plasma membrane H⁺‐ATPase

Two novel effectors of trafficking and maturation of the yeast plasma membrane H⁺‐ATPase

GTPase Sar1 regulates the trafficking and secretion of the virulence factor gp63 in Leishmania

A Functional Study of AUXILIN-LIKE1 and 2, Two Putative Clathrin Uncoating Factors in Arabidopsis

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Auxilin facilitates membrane traffic in the early secretory pathway

Cited by 16 publications

References 49 publications

Two novel effectors of trafficking and maturation of the yeast plasma membrane H+‐ATPase

Two novel effectors of trafficking and maturation of the yeast plasma membrane H+‐ATPase

GTPase Sar1 regulates the trafficking and secretion of the virulence factor gp63 in Leishmania

A Functional Study of AUXILIN-LIKE1 and 2, Two Putative Clathrin Uncoating Factors in Arabidopsis

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Two novel effectors of trafficking and maturation of the yeast plasma membrane H⁺‐ATPase

Two novel effectors of trafficking and maturation of the yeast plasma membrane H⁺‐ATPase