Abstract 1837 ID please! Determining protein interactors of an uncharacterised yeast lipase using BioID
Bibliographic record
Abstract
Drl1 is a putative lipase in Saccharomyces cerevisiae for which the exact role in the lipid metabolic pathways is still unknown. A previous genome-wide screening from our lab showed that Drl1 is involved in the regulation of diacylglycerol (DAG) localization, as normal DAG pools were disrupted in Drl1 knockout and catalytically dead strains. We aim to determine the role of Drl1 in lipid metabolism by studying its protein-protein interactions using TurboID, a more recent iteration of the BioID biotin proximity labelling technology, combined with an auxin-induced degron-fused Bpl1 to reduce endogenous biotinylation signal (https://doi.org/10.15252/msb.202211084). Streptavidin-HRP blots have shown that overnight incubation in auxin-enriched media greatly reduces signal from abundant endogenously biotinylated proteins, including Acc1, Hfa1 and Arc1. Furthermore, four-hour pulses of biotin combined with continued auxin incubation resulted in the appearance of additional bands on Streptavidin-HRP blots compared to controls, providing strong evidence that interactors of Drl1 are being biotinylated above background levels and would make promising candidates for Streptavidin-Sepharose affinity purification, with the end goal of sending enriched samples for proteomic analysis to identify likely interactors of Drl1. Different washes and elution conditions varying pH and denaturing agents, are currently being tested using Streptavidin-coated magnetic beads to determine optimum enrichment conditions for MS-MS analysis. This work has been financially supported by the Natural Sciences and Engineering Research Council of Canada (NSERC) grant to VZ.
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How this classification was reachedexpand
Full frame distilled prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. Learned from the 10,348 direct Codex labels and 10,348 direct Gemma labels. Candidate is the union of thresholded teacher heads; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels or direct frontier model labels.
Codex and Gemma teacher scores by category
| 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 | 0.000 |
| Insufficient payload (model declined to judge) | 0.000 | 0.000 |
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 teacher head, 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".