TgATG9 is required for autophagosome biogenesis and maintenance of chronic infection in Toxoplasma gondii DOI Creative Commons
Pariyamon Thaprawat, Zhihai Zhang,

Eric C. Rentchler

et al.

Autophagy Reports, Journal Year: 2024, Volume and Issue: 3(1)

Published: Oct. 23, 2024

is a ubiquitous protozoan parasite that can reside long-term within hosts as intracellular tissue cysts comprised of chronic stage bradyzoites. To perturb infection requires better understanding the cellular processes mediate persistence. Macroautophagy/autophagy catabolic and homeostatic pathway required for

Language: Английский

Growing thin — How bulk lipid transport drives expansion of the autophagosome membrane but not of its lumen DOI Creative Commons
Thomas J. Melia

Current Opinion in Cell Biology, Journal Year: 2023, Volume and Issue: 83, P. 102190 - 102190

Published: June 27, 2023

Language: Английский

Citations

7
Dynein and dynactin move long-range but are delivered separately to the axon tip DOI Creative Commons
Alexander D. Fellows, Michaela Bruntraeger, Thomas Burgold

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2023, Volume and Issue: unknown

Published: July 3, 2023

Abstract Axonal transport is essential for neuronal survival. This driven by microtubule motors including dynein, which transports cargo from the axon tip back to cell body. function requires its cofactor dynactin and regulators LIS1 NDEL1. Due difficulties imaging dynein at a single-molecule level, it unclear how this motor coordinate along length of axon. Here we use neuron-inducible human stem-celllines (NGN2-OPTi-OX) endogenously tag components visualise them near-single molecule regime. In retrograde direction, find that can move entire (>500 μ m) in one go. Furthermore, NDEL1 also undergo longdistance movement, despite being mainly implicated with initiation transport. Intriguingly, anterograde dynein/LIS1 faster than dynactin/NDEL1 consistent on different cargos. Therefore, neurons ensure efficient holding dynein/dynactin cargos over long distances, but keeping separate until required.

Language: Английский

Citations

7
Rapid in-EPON CLEM: Combining fast and efficient labeling of self-labeling enzyme tags with EM-resistant Janelia Fluor dyes and StayGold DOI Creative Commons
Rico Franzkoch,

Sabrina Wilkening,

Viktoria Liss

et al.

Heliyon, Journal Year: 2024, Volume and Issue: 10(7), P. e28055 - e28055

Published: March 18, 2024

Correlative light and electron microscopy (CLEM) combines (LM) of fluorescent samples to ultrastructural analyses by (EM). Pre-embedding CLEM often suffers from inaccurate correlation between LM EM modalities. Post-embedding enables precise registration structures directly on sections, but requires markers withstanding sample preparation, especially osmium tetroxide fixation, dehydration EPON embedding. Most proteins (FPs) lose their fluorescence during such conventional embedding (CE), synthetic dyes represent promising alternatives as stability exceeds those FP. We analyzed various Janelia Fluor TMR conjugated ligands for self-labeling enzymes, HaloTag, preservation after CE. show that TMR, JF525, JF549, JFX549 JFX554 retain fluorescence, with yielding best results overall, also allowing integration high-pressure freezing freeze substitution. Furthermore, we found the recently published FP StayGold resist CE, facilitating dual-fluorescence in-resin CLEM.

Language: Английский

Citations

2
Mutations in the non-catalytic polyproline motif destabilize TREX1 and amplify cGAS-STING signaling DOI
Abraham Shim, Xiaohan Luan, Wen Zhou

et al.

Human Molecular Genetics, Journal Year: 2024, Volume and Issue: 33(18), P. 1555 - 1566

Published: May 25, 2024

Abstract The cGAS-STING pathway detects cytosolic DNA and activates a signaling cascade that results in type I interferon (IFN) response. endoplasmic reticulum (ER)-associated exonuclease TREX1 suppresses by eliminating from the cytosol. Mutations compromise function are linked to autoinflammatory disorders, including systemic lupus erythematosus (SLE) Aicardi-Goutières syndrome (AGS). Despite key roles regulating suppressing excessive inflammation, impact of many disease-associated mutations—particularly those outside core catalytic domains—remains poorly understood. Here, we characterize recessive AGS-linked P61Q mutation occurring within characterized polyproline helix (PPII) motif. In keeping with its position or ER targeting motifs, neither mutation, nor aggregate proline-to-alanine PPII disrupts activity, subcellular localization, regulation overexpression systems. Introducing targeted mutations into endogenous locus revealed destabilize protein, resulting impaired activity unrestrained activation. Overall, these demonstrate mutations, P61Q, impair proper immune lead autoimmune disease through destabilization.

Language: Английский

Citations

2
TgATG9 is required for autophagosome biogenesis and maintenance of chronic infection in Toxoplasma gondii DOI Creative Commons
Pariyamon Thaprawat, Zhihai Zhang,

Eric C. Rentchler

et al.

Autophagy Reports, Journal Year: 2024, Volume and Issue: 3(1)

Published: Oct. 23, 2024

is a ubiquitous protozoan parasite that can reside long-term within hosts as intracellular tissue cysts comprised of chronic stage bradyzoites. To perturb infection requires better understanding the cellular processes mediate persistence. Macroautophagy/autophagy catabolic and homeostatic pathway required for

Language: Английский

Citations

2