Phosphorylation and binding partners of the human NFE2L3 (NRF3) transcription factor
Bibliographic record
Abstract
Changes in gene expression underlie nearly all biological processes and are governed by the action of transcription factors. Here, we analyzed the NFE2L3 (Nuclear factor (erythroid-derived 2)-like 3) protein, also known as NRF3, a member of the Cap'n'Collar (CNC) transcription factor family. In order to characterize the phosphorylation status of NFE2L3, we created a GST-tagged NFE2L3 fusion protein, which was used as a substrate in a kinase screen developed by Screaton and colleagues (University of Ottawa). Out of 420 kinases assessed, 5 were found to phosphorylate NFE2L3 including polo-like kinase 1 (PLK1). Phosphorylation of NFE2L3 by PLK1 was further confirmed by in vitro kinase assays. Immunoprecipitation studies suggested NFE2L3 interacts with PLK1.Next, we were interested in determining whether glycogen synthase kinase 3 (GSK3) regulates NFE2L3 turnover. Recently, our laboratory discovered NFE2L3 to be ubiquitinated and dependent on the f-box and WD repeat domain-containing 7 (FBW7) ubiquitin ligase (Kannan et al., in preparation). This suggested that GSK3 may be regulating these events as it has been documented extensively to function in concert with FBW7 by first phosphorylating its substrates thereby facilitating FBW7 binding. Corroborating with these data, we showed that GSK3 can bind and phosphorylate NFE2L3. We also found that NFE2L3 expression was stabilized using an inhibitor of GSK3 as well as upon knockdown of GSK3. Finally, GSK3 knockdown rescued FBW7 driven destabilization of NFE2L3. This showed that FBW7-mediated turnover of NFE2L3 is dependent on GSK3 phosphorylation. The final goal of this project was to identify novel NFE2L3 interacting proteins. In order to do so, immunoprecipitation together with mass spectrometric analysis was utilized. These experiments uncovered a series of potential interactors. Of these, the protein golgin-84 was shown to be a bona fide binding partner of NFE2L3. These data indicated the transcription factor may be targeted to the golgi.
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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 | 0.001 | 0.000 |
| Meta-epidemiology (narrow) | 0.001 | 0.000 |
| Meta-epidemiology (broad) | 0.001 | 0.000 |
| Bibliometrics | 0.000 | 0.001 |
| Science and technology studies | 0.001 | 0.000 |
| Scholarly communication | 0.000 | 0.002 |
| Open science | 0.002 | 0.000 |
| Research integrity | 0.001 | 0.001 |
| 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".