Dynamic trafficking and turnover of Jam-C is essential for endothelial cell migration

Kostelnik, Katja B.; Barker, Amy R.; Schultz, Christopher; Rajeeve, Vinothini; White, Ian J.; Aurrand-Lions, Michel; Nourshargh, Sussan; Cutillas, Pedro R.; Nightingale, Thomas D.

doi:10.1101/625913

“…VEGF stimulation increased the cell-surface associated JAM-C fraction to 60 percent in approximately one hour. A more recent study on JAM-C dynamics in quiescent HUVECs reported that two thirds of the cell-surface JAM-C fraction was removed from the cell junctions in two hours [110]. The cytoplasmic JAM-C was detected partially in early endosomes and in MVBs, indicating that it underwent lysosomal degradation.…”

Section: Junctional Adhesion Molecule (Jam)mentioning

confidence: 92%

“…The internalized proteins subsequently collocated with markers of early endosomes and with Rab4 and -11, markers of 'fast' and 'slow' recycling, respectively [23]. Either calcium chelation or stimulation with TNFα induced endocytosis of JAM-C in HUVECs [110]. Subsequently, it was observed in tubular extensions from membranous structures formed at the cell junctions, but the endocytic pathway was not identified.…”

Section: Junctional Adhesion Molecule (Jam)mentioning

confidence: 99%

Membrane Trafficking of Integral Cell Junction Proteins

Horowitz¹

2021

Preprint

0

View full text Add to dashboard Cite

Though membrane trafficking of cell junction proteins has been studied extensively for more than two decades, the accumulated knowledge remains fragmentary. The goal of this review is to synthesize the numerous and disparate observations on the membrane trafficking of the five major junction transmembrane proteins accumulated to date to comprehend the current state of the art, to highlight differences and similarities among the trafficking pathways, and to identify topics that are not fully understood. Clathrin-mediated endocytosis appears to be the main but not exclusive mode of internalization. Caveolin-mediated endocytosis and macropinocytosis are employed less frequently. PDZ-domain binding is the predominant mode of interaction between junction protein cytoplasmic tails and scaffold proteins. It is shared by claudins, the largest family of junction integral proteins, and by the three nectins. All five proteins are destined to either recycling via Rab4/Rab11 GTPases or to degradation. The sorting mechanisms that underlie the specificity of their endocytic pathways and determine their fates are not fully known. New data is presented to introduce an emerging role of junction-associated scaffold proteins in claudin membrane trafficking.

show abstract

“…Variable modifications included in searches were oxidation of methionine, pyro-glu (N-term) and phosphorylation of serine, threonine and tyrosine. The mascot result (DAT) files were extracted into Excel files for further normalisation, quantitative label-free analysis and statistical analysis as described previously (49,96).…”

Section: Mass Spectrometrymentioning

confidence: 99%

“…Consistent with this, the full repertoire of proteins interacting with LC3 is not known. Here we systematically defined the LC3B interactome by performing an enzymatic proximity tagging approach using an engineered ascorbate peroxidase, APEX2 (48,49). We used HeLa cells stably transfected with an APEX2-and GFPtagged LC3B construct (or the control GFP-LC3B) and cultured them in normal growth conditions.…”

Section: Introductionmentioning

confidence: 99%

Proximity interactome of LC3B in normal growth conditions

Nollet

¹

,

Agrotis

²

,

Michailidis

³

et al. 2021

Preprint

Self Cite

0

View full text Add to dashboard Cite

LC3 (Light Chain 3) is a key player of autophagy, a major stress-responsive proteolysis pathway promoting cellular homeostasis. It coordinates the formation and maturation of autophagosomes and recruits cargo to be further degraded upon autophagosome-lysosome fusion. To orchestrate its functions, LC3 binds to multiple proteins from the autophagosomes inner and outer membranes, but the full extent of these interactions is not known. Moreover, LC3 has been increasingly reported in other cellular locations than the autophagosome, with cellular outcome not fully understood and not all related to autophagy. Furthermore, novel functions of LC3 as well as autophagy can occur in cells growing in a normal medium thus in non-stressed conditions. A better knowledge of the molecule in proximity to LC3 in normal growth conditions will improve the understanding of LC3 function in autophagy and in other cell biology function. Using an APEX2 based proteomic approach, we have detected 407 proteins in proximity to the well-characterised LC3B isoform in non-stress conditions. These include known and novel LC3B proximity proteins, associated with various cell localisation and biological functions. Sixty-nine of these proteins contain a putative LIR (LC3 Interacting Region) including 41 not reported associated to autophagy. Several APEX2 hits were validated by co-immunoprecipitation and co-immunofluorescence. This study uncovers the LC3B global interactome and reveals novel LC3B interactors, irrespective of LC3B localisation and function. This knowledge could be exploited to better understand the role of LC3B in autophagy and non-autophagy cellular processes.

show abstract

“…The internalized proteins subsequently collocated with markers of early endosomes and with Rab4 and -11, markers of 'fast' and 'slow' recycling, respectively [23]. Either calcium chelation or stimulation with TNFα induced endocytosis of JAM-C in HUVECs [117]. Subsequently, it was observed in tubular extensions from membranous structures formed at the cell junctions, but the endocytic pathway was not identified.…”

Section: Junctional Adhesion Molecule (Jam)mentioning

confidence: 99%

Membrane Trafficking of Integral Cell Junction Proteins and its Functional Consequences

Horowitz¹

2021

Preprint

0

View full text Add to dashboard Cite

Though membrane trafficking of cell junction proteins has been studied extensively for more than two decades, the accumulated knowledge remains fragmentary. The goal of this review is to synthesize the numerous and disparate observations on the membrane trafficking of the five major junction transmembrane proteins accumulated to date to comprehend the current state of the art, to highlight differences and similarities among the trafficking pathways, and to identify topics that are not fully understood. Clathrin-mediated endocytosis appears to be the main but not exclusive mode of internalization. Caveolin-mediated endocytosis and macropinocytosis are employed less frequently. PDZ-domain binding is the predominant mode of interaction between junction protein cytoplasmic tails and scaffold proteins. It is shared by claudins, the largest family of junction integral proteins, and by the three nectins. All five proteins are destined to either recycling via Rab4/Rab11 GTPases or to degradation. The sorting mechanisms that underlie the specificity of their endocytic pathways and determine their fates are not fully known. New data is presented to introduce an emerging role of junction-associated scaffold proteins in claudin membrane trafficking.

show abstract

Dynamic trafficking and turnover of Jam-C is essential for endothelial cell migration

Abstract: 15

Cited by 5 publications

References 65 publications

Membrane Trafficking of Integral Cell Junction Proteins

Membrane Trafficking of Integral Cell Junction Proteins

Proximity interactome of LC3B in normal growth conditions

Membrane Trafficking of Integral Cell Junction Proteins and its Functional Consequences

Contact Info

Product

Resources

About