Penta-EF-Hand Protein Peflin Is a Negative Regulator of ER-To-Golgi Transport

Rayl, Mariah; Truitt, Mishana; Held, Aaron; Sargeant, John; Thorsen, Kevin; Hay, Jesse

doi:10.1371/journal.pone.0157227

Cited by 14 publications

(43 citation statements)

References 39 publications

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“…During the preparation of this paper, Rayl et al . reported that peflin is a negative regulator of ER‐to‐Golgi transport by suppressing the interaction between ALG‐2 and Sec31A. In the present study, we found several proteins that interact Ca 2+ ‐dependently with the dimer of FLAG‐ALG‐2 E47A/E114A /ALG‐2 by immunoprecipitation with anti‐FLAG antibody (Fig.…”

Section: Discussionsupporting

confidence: 63%

“…Although there is accumulating evidence indicating that ALG-2 interacts with various proteins [35], there have been only a few reports of proteins interacting with peflin [36,37]. During the preparation of this paper, Rayl et al [38] reported that peflin is a negative regulator of ER-to-Golgi transport by suppressing the interaction between ALG-2 and Sec31A. In the present study, we found several proteins that interact Ca 2+ -dependently with the dimer of FLAG-ALG-2 E47A/E114A /ALG-2 by immunoprecipitation with anti-FLAG antibody (Fig.…”

Section: Discussionmentioning

confidence: 99%

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The calcium‐binding protein ALG‐2 promotes endoplasmic reticulum exit site localization and polymerization of Trk‐fused gene (TFG) protein

et al. 2016

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Apoptosis‐linked gene 2 (ALG‐2), which is a gene product of PDCD6, is a 22‐kDa Ca2+‐binding protein. Accumulating evidence points to a role for ALG‐2 as a Ca2+‐responsive adaptor protein. On binding to Ca2+, ALG‐2 undergoes a conformational change that facilitates its interaction with various proteins. It also forms a homodimer and heterodimer with peflin, a paralog of ALG‐2. However, the differences in cellular roles for the ALG‐2 homodimer and ALG‐2/peflin heterodimer are unclear. In the present study, we found that Trk‐fused gene (TFG) protein interacted with the ALG‐2 homodimer. Immunostaining analysis revealed that TFG and ALG‐2 partially overlapped at endoplasmic reticulum exit sites (ERES), a platform for COPII‐mediated protein transport from the endoplasmic reticulum. Time‐lapse live‐cell imaging demonstrated that both green fluorescent protein‐fused TFG and mCherry‐fused ALG‐2 are recruited to ERES after thapsigargin treatment, which raises intracellular Ca2+ levels. Furthermore, overexpression of ALG‐2 induced the accumulation of TFG at ERES. TFG has an ALG‐2‐binding motif and deletion of the motif decreased TFG binding to ALG‐2 and shortened its half‐life at ERES, suggesting a critical role for ALG‐2 in retaining TFG at ERES. We also demonstrated, by in vitro cross‐linking assays, that ALG‐2 promoted the polymerization of TFG in a Ca2+‐dependent manner. Collectively, the results suggest that ALG‐2 acts as a Ca2+‐sensitive adaptor to concentrate and polymerize TFG at ERES, supporting a potential role for ALG‐2 in COPII‐dependent trafficking from the endoplasmic reticulum.

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Section: Discussionsupporting

confidence: 63%

Section: Discussionmentioning

confidence: 99%

The calcium‐binding protein ALG‐2 promotes endoplasmic reticulum exit site localization and polymerization of Trk‐fused gene (TFG) protein

et al. 2016

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“…For example, most in vitro transport reconstitutions and results with purified ALG-2 have indicated that the protein is an inhibitor of vesicle budding or fusion (Bentley et al, 2010;la Cour et al, 2013). On the other hand some recent intact cell trafficking experiments indicate a suppressive role for ALG-2 based upon ALG-2 depletion (Shibata et al, 2015), while we implied a stimulatory role for ALG-2 because peflin suppressed transport by antagonizing stimulatory ALG-2-Sec31A interactions (Rayl et al, 2016).…”

Section: Introductionmentioning

confidence: 64%

“…The PEF protein apoptosis-linked gene-2 (ALG-2) acts as a Ca 2+ sensor at ER exit sites and stabilizes association of sec31 with the membrane when Ca 2+ is present (Yamasaki et al, 2006;la Cour et al, 2007;Shibata et al, 2007;. Most ALG-2 in cell extracts exists in a stable heterodimer with the PEF protein peflin that binds ALG-2 in a Ca 2+ -inhibited manner (Kitaura et al, 2001; and has been shown to suppress ER export of the cargo marker VSVG-GFP, perhaps by modulating ALG-2 availability to bind ERES (Rayl et al, 2016). Despite all of these observations, a unified model for when and how PEF proteins modulate secretion has not emerged.…”

Section: Introductionmentioning

confidence: 99%

ALG-2 and Peflin Stimulate or Inffibit Copii Targeting and Secretion in Response to Calcium Signaling

Sargeant

Seiler

Costain

et al. 2020

Preprint

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Penta EF-hand (PEF) proteins apoptosis-linked gene 2 (ALG-2) and peflin are cytoplasmic Ca 2+ sensors for which physiological roles are still emerging. Here we demonstrate that adjustment of the ALG-2:peflin expression ratio can modulate basal ER export rates up or down by 107% of the basal rate. Through their ALG-2 subunit, ALG-2-peflin hetero-oligomers are shown to bind ERES and inhibit ER export of multiple cargo types, including collagen I, while ALG-2 by itself binds ERES to stimulate cargo sorting and ER export. During sustained Ca 2+ signaling, peflin and ALG-2 are demonstrated to sharply lower ER export rates by 40% in a novel secretory response to demanding physiological conditions. However, in highly Ca 2+stressed cells, peflin's suppressive role appears to promote pro-apoptotic unfolded protein response (UPR) signaling, indicating that the regulation is not necessarily pro-survival. Thus, regulation of secretion by PEF protein sub-complex binding to ERES in response to Ca 2+ signals impacts key cellular decision processes relevant to aging, neurodegeneration and diabetes.

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“…3A]. There is also evidence that PEF1 and ALG-2 together control transport of smaller cargoes such as the temperature sensitive viral glycoprotein VSV-G (Rayl et al 2016). VSV-G has been used for many years as a reporter of biosynthetic membrane trafficking (Gallione and Rose 1985) and is widely considered a marker of canonical ER-to-Golgi-to-plasma membrane transport.…”

Section: Flexibility Of the Copii Coat: The Case For Large Copii-depementioning

confidence: 99%

COPII-dependent ER export in animal cells: adaptation and control for diverse cargo

McCaughey

Stephens

2018

Histochem Cell Biol

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The export of newly synthesized proteins from the endoplasmic reticulum is fundamental to the ongoing maintenance of cell and tissue structure and function. After co-translational translocation into the ER, proteins destined for downstream intracellular compartments or secretion from the cell are sorted and packaged into transport vesicles by the COPII coat protein complex. The fundamental discovery and characterization of the pathway has now been augmented by a greater understanding of the role of COPII in diverse aspects of cell function. We now have a deep understanding of how COPII contributes to the trafficking of diverse cargoes including extracellular matrix molecules, developmental signalling proteins, and key metabolic factors such as lipoproteins. Structural and functional studies have shown that the COPII coat is both highly flexible and subject to multiple modes of regulation. This has led to new discoveries defining roles of COPII in development, autophagy, and tissue organization. Many of these newly emerging features of the canonical COPII pathway are placed in a context of procollagen secretion because of the fundamental interest in how a coat complex that typically generates 80-nm transport vesicles can package a cargo reported to be over 300 nm. Here we review the current understanding of COPII and assess the current consensus on its role in packaging diverse cargo proteins.

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Penta-EF-Hand Protein Peflin Is a Negative Regulator of ER-To-Golgi Transport

Cited by 14 publications

References 39 publications

The calcium‐binding protein ALG‐2 promotes endoplasmic reticulum exit site localization and polymerization of Trk‐fused gene (TFG) protein

The calcium‐binding protein ALG‐2 promotes endoplasmic reticulum exit site localization and polymerization of Trk‐fused gene (TFG) protein

ALG-2 and Peflin Stimulate or Inffibit Copii Targeting and Secretion in Response to Calcium Signaling

COPII-dependent ER export in animal cells: adaptation and control for diverse cargo

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