High Temporal Resolution Reveals Simultaneous Plasma Membrane Recruitment of TPLATE Complex Subunits

Wang, Jie; Mylle, Evelien; Johnson, Alexander; Besbrugge, Nienke; Jaeger, Geert De; Friml, Jiřı́; Pleskot, Roman; Damme, Daniël Van

doi:10.1104/pp.20.00178

Cited by 30 publications

(48 citation statements)

References 51 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The AtEH/Pan1 proteins are more loosely associated with TPC as they for example do not associate with the other complex subunits when a truncated TML subunit forces TPC into the cytoplasm (Gadeyne et al, 2014) and they do not co-purify with the complex in Dictyostelium (Hirst et al, 2014). Recent evidence suggests however a similar arrival time of all TPC subunits at the plasma membrane at the onset of endocytosis, preceding clathrin arrival (Gadeyne et al, 2014;Narasimhan et al, 2020;Wang et al, 2020). The AtEH/Pan1 proteins are the plant homologs of yeast Pan1p, which is known for its role as an activator of ARP2/3-dependent actin dynamics during endocytosis (Duncan et al, 2001;Toshima et al, 2005Toshima et al, , 2007.…”

Section: Introductionmentioning

confidence: 99%

Distinct EH domains of the endocytic TPLATE complex confer lipid and protein binding

Yperman

Papageorgiou

Merceron

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

Clathrin-mediated endocytosis (CME) is the gatekeeper of the plasma membrane. In contrast to animals and yeasts, CME in plants depends on the TPLATE complex (TPC), an evolutionary ancient adaptor complex. The mechanistic contribution of the individual TPC subunits to plant CME remains however elusive. In this study, we used a multidisciplinary approach to elucidate the structural and functional roles of the evolutionary conserved N-terminal Eps15 homology (EH) domains of the TPC subunit AtEH1/Pan1. By integrating high-resolution structural information obtained by X-ray crystallography and NMR spectroscopy with all-atom molecular dynamics simulations, we provide structural insight into the function of both EH domains. Whereas one EH domain binds negatively charged PI(4,5)P2 lipids, unbiased peptidome profiling by mass-spectrometry revealed that the other EH domain interacts with the double N-terminal NPF motif of a novel TPC interactor, the integral membrane protein Secretory Carrier Membrane Protein 5 (SCAMP5). Furthermore, we show that AtEH/Pan1 proteins control the internalization of SCAMP5 via this double NPF peptide interaction motif. Collectively, our structural and functional studies reveal distinct but complementary roles of the EH domains of AtEH/Pan1 have in plant CME and connect the internalization of SCAMP5 to the TPLATE complex.

show abstract

Section: Introductionmentioning

confidence: 99%

Distinct EH domains of the endocytic TPLATE complex confer lipid and protein binding

Yperman

Papageorgiou

Merceron

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…As previously hypothesized, concomitantly with this loss in Dictyostelium, the TCUP subunit lost some of its functions and the connection to other subunits ( 6 ). A tight interaction between TML and the AtEH/Pan1 proteins is present as high temporal resolution spinning disc data shows an identical recruitment to the plasma membrane of AtEH proteins and the core subunits of TPC ( 22 ). This complements the data obtained in this study via Y3H and in planta protein-protein interaction assays that TML associates with the AtEH/Pan1 proteins (Fig.…”

Section: Resultsmentioning

confidence: 93%

“…μHDs are a common feature among vesicle trafficking complexes and are not only known to be involved in both accessory proteins and cargo interactions but have also been shown to directly interact with lipid membranes (Dergai et al, 2010; Henne et al, 2010; Jackson et al, 2012; Reider et al, 2009; Shimada et al, 2016). An unequal expression of TML and TPLATE in a double complemented, double mutant Arabidopsis line resulted in the dynamic recruitment of TML to the plasma membrane without the presence of TPLATE but not the other way around (Wang et al, 2020). Therefore, we hypothesized that TML μHD might provide simultaneous membrane recruitment and association with the AtEH/Pan1 subunits.…”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

The TPLATE subunit is essential for structural assembly of the endocytic TSET complex

Yperman

Wang

Eeckhout

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

All eukaryotic cells rely on endocytosis to regulate the plasma membrane proteome and lipidome. Most eukaryotic groups, with the exception of fungi and animals, have retained the evolutionary ancient TSET complex as a regulator of endocytosis. Despite the presence of similar building blocks in TSET, compared to other coatomer complexes, structural insight into this adaptor complex is lacking. Here, we elucidate the molecular architecture of the octameric plant TSET complex (TPLATE complex/TPC) using an integrative structural approach. This allowed us to describe a plant-specific connection between the TML subunit and the AtEH/Pan1 proteins and show a direct interaction between the complex and the plasma membrane without the need for any additional protein factors. Furthermore, we identify the appendage of TPLATE as crucial for complex assembly. Structural elucidation of this ancient adaptor complex vastly advances our functional as well as evolutionary insight into the process of endocytosis.

show abstract

“…1E). Very recently, the temporal resolution of endocytic spinning disc microscopy imaging in Arabidopsis was increased by lowering the temperature of the samples (Wang et al ., 2020).…”

Section: Live‐cell Visualisation Of Endocytic Internalisationmentioning

confidence: 99%

Visualising endocytosis in plants: past, present, and future

Dragwidge

Damme

2020

Journal of Microscopy

Self Cite

View full text Add to dashboard Cite

Chris Hawes had a lively fascination for the immensely complex organisation of the endomembrane system, including the process of endocytosis. This is the method by which eukaryotic cells internalise membrane proteins, lipids, carbohydrates, and cell wall enzymes from the cell surface through membrane bound vesicles. Endocytosis occurs progressively, starting with early membrane deformation, scission, and finally the release of the vesicle into the cytoplasm. Next to secretion, endocytosis allows the cell to control the proteome composition of its inner and outer surface membrane and as such, its communication with the outside world. Whereas endocytosis was initially considered theoretically impossible in plants due to their high turgor pressure, it is now established as essential for plant life. Furthermore, endocytosis remains a highly active field of research, both in yeast, animal, and plant model systems. Over the past three decades, the tools and techniques used to visualise, quantify, and characterise endocytosis have resulted in an increasingly higher spatiotemporal understanding of this process. Here we provide a brief history of plant endocytosis research from the time when Chris Hawes was investigating the process, to the current state-of-the-art in the field. We will end this chapter with a discussion on some promising future developments for plant endocytosis research. A brief history of endocytosis in plants Chris Hawes had an intense fascination for understanding the intracellular organisation of a cell. Specifically, Chris was intrigued by the immensely complex organisation of the endomembrane system, which covers the anterograde trafficking pathways from the endoplasmic reticulum to the Golgi apparatus, to the trans-Golgi network and to the plasma membrane, as well as the retrograde transport route, which recycles material from the Golgi apparatus back to the endoplasmic reticulum. Moreover, he also studied the endocytic

show abstract

High Temporal Resolution Reveals Simultaneous Plasma Membrane Recruitment of TPLATE Complex Subunits

Abstract: Lowering the temperature increases the temporal resolution of visualizing protein recruitment at the plasma membrane, demonstrated by dual-color live cell imaging of endocytic complex subunits.

Cited by 30 publications

References 51 publications

Distinct EH domains of the endocytic TPLATE complex confer lipid and protein binding

Distinct EH domains of the endocytic TPLATE complex confer lipid and protein binding

The TPLATE subunit is essential for structural assembly of the endocytic TSET complex

Visualising endocytosis in plants: past, present, and future

Contact Info

Product

Resources

About