Biochemical Analysis of Wsc1p and Mid2p Interacting Partners under Thermal Stress
Bibliographic record
Abstract
The PKC1 cell wall integrity pathway (CWIP) is essential for survival and maintenance of cell wall integrity in yeast cells. Upon exposure to cell wall stress, signaling proteins (sensors) in the plasma membrane activate this pathway by interacting with other signaling proteins (interactors). The objective of this research is to know if interactors previously identified by the integrated Membrane Yeast Two‐Hybrid (iMYTH) technique contribute to CWIP activation under thermal stress. Our hypothesis is that interactors of the Wsc1p and Mid2p stress sensors are required for survival to thermal stress. To test this hypothesis, strains bearing null mutations for genes encoding stress sensor proteins and their interacting protein partners were tested for CWIP activation at 37°C by Western Blot technique. The cells were exposed 1hr at 37°C during log phase and the protein extracts were tested with antibodies that measure CWIP activation through Slt2p phosphorylation. Most of the null mutants showed normal PKC1 activation when shifted from growth at 27°C to 37°C similar to a wild type strain. Deletion of GTT1 an interactor of Mid2p exhibited very high levels of Slt2p phosphorylation in both conditions relative to the wild type suggesting that PKC1 was being activated constitutively. In conclusion, interactors of Wsc1p and Mid2p can function as signaling proteins involved in PKC1 pathway regulation and may be associated with survival under different stresses. This research was supported by the UPR‐MSC, Univ. of Toronto, Univ. of Kentucky, NIGMS/NIAID‐SC1AI081658, NCRR‐RCMI‐G12MD007600, MBRS‐RISE‐R25GM061838, and NIH‐INBRE 8 P20 GM103475‐13.
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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 | 0.001 |
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".