Characterization and mutant analysis of the Drosophila melanogaster homologues of amphiphysin and enthoprotin, two proteins implicated in vesicular trafficking
Bibliographic record
Abstract
Vesicular trafficking is necessary in eukaryotic organisms to move proteins and membrane from organelle to organelle or plasma membrane. Budding and scission of vesicles requires the interplay of a number of cytoplasmic proteins, many of which also interact with membranes. The study of such molecules is frequently defined either in biochemical assays or through the expression of various constructs in a cell culture system. What is often missing is insight into function in an intact multicellular organism. Drosophila inelanogaster is an excellent model for studying such proteins as its development is well characterized and it has well-defined genetics to allow for the study of mutants. In this thesis, I have chosen to investigate amphiphysin and enthoprotin, two membrane-interacting proteins implicated in the formation of clathrin-coated vesicles. Amphiphysin contains multiple protein interaction domains and is thought to act as a scaffold during late steps of clathrin-mediated endocytosis particularly at synapses. Based on studies in yeast and vertebrate cell culture, enthoprotin is thought to help form clathrin-coated pits in the trans-Golgi network (TON) to endosome trafficking pathway. It is believed to act by promoting membrane curvature and by binding to both clathrin and clathrin adaptors. Drosophila amphiphysin (Amph) is not found at the pre-synaptic membrane, but rather at the post-synaptic membrane. Amph mutants are viable and have a locomotory defect. However, they show no defects in synaptic neurophysiology, suggesting a role for Amph in muscle development. Drosophila Enthoprotin (Enth) is localized to the TGN and endosome, consistent with its predicted function. Enth mutants are viable with slightly roughened eyes, but otherwise appear normal. However, oogenesis is disrupted at a late stage and females mutants are completely sterile. Analysis of germline clones shows that Enth is required only in the follicle cells for normal oogenesis. Additionally, Enth mutant clones show a cell growth phenotype that appears to be due to perturbation of the Akt pathway. Amph functions quite differently in Drosophila than work from vertebrate cell culture had suggested. Enth seems to function where expected, but mutant analysis revealed unexpected phenotypes. Together, this work highlights the value of studying proteins in vivo.
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How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.
Distilled classifier scores by category (both heads)
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.000 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.001 |
| Insufficient payload (model declined to judge) | 0.001 | 0.001 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".