Asymmetric organelle positioning during epithelial polarization of C. elegans intestinal cells

Brandt, James N; Voss, Laura; Rambo, Fiona M.; Nicholson, Katelyn; Thein, Jackson R.; Fairchild, Lydia; Seabrook, Laurence J.; Lewis, Danielia; Guevara-Hernandez, Lali; White, Matthew L.; Sax, Luca; Eichten, Victoria; Harper, Logan R.; Hermann, Greg J.

doi:10.1016/j.ydbio.2021.09.007

Cited by 8 publications

(4 citation statements)

References 96 publications

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“…6A ; proposed polarity model; compare ( 13 )], the apical directionality of biosynthetic vesicles should depend on bcAMs and actin during de novo membrane biogenesis and polarity establishment. The polarization of vesicles (and nuclei) toward the future apical domain was indeed noted to predate membrane polarity establishment in the C. elegans intestine ( 26 , 28 ). It remains difficult in any in vitro or in vivo system to visually track the directionality of biosynthetic trafficking, since polarized cargo becomes successively enriched from the endoplasmic reticulum (ER) to the plasma membrane, while vesicle-membrane and coat components are consecutively recruited and shed ( 29 ).…”

Section: Resultsmentioning

confidence: 99%

Branched-chain actin dynamics polarizes vesicle trajectories and partitions apicobasal epithelial membrane domains

et al. 2023

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In prevailing epithelial polarity models, membrane- and junction-based polarity cues such as the partitioning-defective PARs specify the positions of apicobasal membrane domains. Recent findings indicate, however, that intracellular vesicular trafficking can determine the position of the apical domain, upstream of membrane-based polarity cues. These findings raise the question of how vesicular trafficking becomes polarized independent of apicobasal target membrane domains. Here, we show that the apical directionality of vesicle trajectories depends on actin dynamics during de novo polarized membrane biogenesis in the C. elegans intestine. We find that actin, powered by branched-chain actin modulators, determines the polarized distribution of apical membrane components, PARs, and itself. Using photomodulation, we demonstrate that F-actin travels through the cytoplasm and along the cortex toward the future apical domain. Our findings support an alternative polarity model where actin-directed trafficking asymmetrically inserts the nascent apical domain into the growing epithelial membrane to partition apicobasal membrane domains.

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Section: Resultsmentioning

confidence: 99%

Branched-chain actin dynamics polarizes vesicle trajectories and partitions apicobasal epithelial membrane domains

et al. 2023

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“…If bcAMs/actin were to specify cellular membrane polarity by asymmetrically inserting newly-synthesized apical membrane components into the expanding membrane (proposed model; Fig.6a), the apical directionality of vesicles should depend on bcAMs/actin at the time of de novo polarized membrane biogenesis. Indeed, vesicular (and nuclear) repositioning has been noted to occur during the establishment of C. elegans intestinal polarity and its contribution to polarity establishment has been hypothesized but not yet demonstrated 23, 24 . The visual tracking of biosynthetic vesicles from the ER to the plasma membrane in single cells has been difficult in any system, since biosynthetic cargo is thought to be successively enriched on its way to the plasma membrane while vesicle-coat and vesicle-membrane components are consecutively shed and recruited on this route 25, 26 .…”

Section: Main Textmentioning

confidence: 99%

A transcytotic actin shift polarizes vesicle trajectories and partitions apicobasal epithelial membrane domains

Jafari

Khan

Zhang

et al. 2022

Preprint

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In prevailing epithelial polarity models, membrane-based polarity cues such as the partitioning-defective PARs specify the positions and identities of apicobasal membrane domains. Recent findings suggest, however, that vesicle-associated polarity cues specify membrane polarity by positioning the apical domain, upstream of membrane-based polarity cues. These findings raised the question how vesicles acquire apicobasal directionality independent of polarized target membrane domains. Here, we show that the apical directionality of vesicle trajectories depends on intracellular actin dynamics during the establishment of membrane polarity in the C. elegans intestine. We find that actin, powered by branched-chain actin dynamics, determines the position of apical membrane components, PARs, and itself on expanding membranes. Using photomodulation, we demonstrate that F-actin travels through the cytoplasm and along the cortex towards the future apical domain. Our findings suggest an alternative polarity model where actin-dependent directional trafficking inserts the nascent apical domain into the growing membrane to partition its apicobasal domains.

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“…These are: PGP-2, a member of the ABC family of transmembrane transporters (24); CDF-2, a zinc transporter (6); and GLO-3, a nematode membrane protein (57). Of these three, both PGP-2 and CDF-2 have been used as markers of gut granules (26, 58, 59), and can be considered the best -characterized examples of proteins that localize to the gut granules. Comparing the proteomes found in 11 intestinal samples from individual glo-1(-) animals to those found in wild-type intestinal tissue samples, we identified GLO-1 and 22 other proteins that were largely missing in glo-1 mutant tissue and so are likely part of the gut granule proteome (Fig.…”

Section: Resultsmentioning

confidence: 99%

Single tissue proteomics inCaenorhabditis elegansreveals proteins resident in intestinal lysosome-related organelles

Tan,

Wang,

Park

et al. 2023

Preprint

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The nematode intestine is the primary site for nutrient uptake and storage as well as the synthesis of biomolecules; lysosome-related organelles known as gut granules are important for many of these functions. Aspects of intestine biology are not well understood, including the export of the nutrients it imports and the molecules it synthesizes, as well as the complete functions and protein content of the gut granules. Here, we report a mass spectrometry-based proteomic analysis of the intestine of theCaenorhabditis elegansand of its gut granules. Overall, we identified approximately 5,000 proteins each in the intestine and the gonad and showed that most of these proteins can be detected in samples extracted from a single worm, suggesting the feasibility of individual-level genetic analysis using proteomes. Comparing proteomes and published transcriptomes of the intestine and the gonad, we identified proteins that appear to be synthesized in the intestine and then transferred to the gonad. To identify gut granule proteins, we compared the proteome of individual intestines deficient in gut granules to the wild-type. The identified gut granule proteome includes proteins known to be exclusively localized to the granules and additional putative novel gut granule proteins. We selected two of these putative gut granule proteins for validation via immunohistochemistry, and our successful confirmation of both suggests that our strategy was effective in identifying the gut granule proteome. Our results demonstrate the practicability of single tissue mass-spectrometry- based proteomic analysis in small organisms and in its utility for making discoveries.Significance statementWe show that tissue-specific proteomic analysis is achievable and can be done efficiently at an individual level in a small nematode, with resolution sufficient for genetic analysis on a single animal basis. With data collected from single animals, we produced high-quality sets of proteins that described the proteomes of the gonad and the intestine. Comparison of these proteomes with the organs’ transcriptomes improved our understanding of interorgan protein transport. We applied single-tissue proteomic to describe the proteome of the gut granules in the nematode intestine, a specialized lysosome-related organelle with important functions but which is not well characterized, identifying proteins not previously known to be associated with LROs and verifying two by subcellular localization.

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Asymmetric organelle positioning during epithelial polarization of C. elegans intestinal cells

Cited by 8 publications

References 96 publications

Branched-chain actin dynamics polarizes vesicle trajectories and partitions apicobasal epithelial membrane domains

Branched-chain actin dynamics polarizes vesicle trajectories and partitions apicobasal epithelial membrane domains

A transcytotic actin shift polarizes vesicle trajectories and partitions apicobasal epithelial membrane domains

Single tissue proteomics inCaenorhabditis elegansreveals proteins resident in intestinal lysosome-related organelles

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