Analysis of novel interacting protein partners of Wsc1p and Mid2p, two sensor proteins of the PKC1 signaling pathway
Bibliographic record
Abstract
Wsc1 and Mid2 are protein sensors of cell wall stress that activate the PKC1‐Cell Wall Integrity Pathway in response to environmental stress. We hypothesized that Wsc1p and Mid2p attract additional interacting protein partners (interactors) that can be associated with PKC1 activation. To identify novel interactors, the integrated membrane Y2H (iMYTH) technique was applied. A cDNA plasmid library of prey proteins was expressed in bait strains containing iMYTH reporter modules of Wsc1p and Mid2p. The interacting prey plasmid clones were recovered and sequenced. To assess their functionality in PKC1 activation, we performed Western blot analysis of phospho‐Slt2p in null mutants of the interactors. Under normal culture conditions at 30°C, 14 novel interactors were confirmed for Wsc1p and 29 for Mid2p. In a second iMYTH screen performed under heat stress conditions at 37°C, 4 novel interactors for Mid2p and 1 for Wsc1p were identified. At 37°C, YHR193C was the dominant interactor for Wsc1p and Mid2p. Null mutants of the interactors YOL109W, YHR135C, YPL199C exhibited normal PKC1 activation status at 30°C and 37°C while theYHR193C null mutant failed to activate PKC1 at 37°C. In conclusion, YHR193C is a protein interactor of Wsc1p and Mid2p required for activation of the PKC1 pathway. Supported by NCRR‐RCMI‐G12RR03051, MBRS‐RISE‐R25GM061838 & Univ. of Toronto.
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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) | 0.001 | 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) | 0.001 | 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 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".