<i>Bacillus subtilis</i> MurJ and Amj Lipid II flippases are not essential for growth
Bibliographic record
Abstract
ABSTRACT Identification of the protein that mediates transbilayer transport of the undecaprenyl-pyrophosphate (Und-PP) linked peptidoglycan precursor Lipid II has long been a subject of investigation. Candidates belonging to both the MOP (multidrug/oligosaccharidyl-lipid/polysaccharide) and SEDS (shape, elongation, division and sporulation) families of transmembrane proteins have been proposed, exhibiting characteristics consistent with mediating this process, including genetic essentiality and biochemical activity. While MOP family proteins including MurJ are widely considered to be the primary Lipid II transporter, questions still remain including a role for the SEDS proteins in this process. We and others previously showed that a Bacillus subtilis strain lacking all four MurJ homologs is viable, thereby implicating a separate mode of Lipid II transport across the membrane. However, a subsequent report of synthetic essentiality between B. subtilis MurJ and the flippase Amj suggested that they are necessary and sufficient. Here, we show that this effect is alleviated by excess synthesis of the enzyme responsible for Und-PP production. Thus, the inviability of a murJ-amj double mutant strain is not due to the essentiality of these enzymes for flipping Lipid II but is instead most likely a consequence of a reduction of free Und-PP levels. This result is consistent with a non-MOP-dependent pathway for Lipid II transport across the cytoplasmic membrane to enable cell wall peptidoglycan synthesis. IMPORTANCE The assembly of peptidoglycan (PG), the typically essential polymer that provides structural integrity to bacterial cells, begins with the synthesis of the Lipid II monomer in the cytoplasm and along the cytoplasmic face of the inner membrane. Lipid II is then translocated across the membrane to the extracellular site of polymerization. The mechanistic basis for this process remains unclear, with genetic and/or biochemical evidence pointing to two different families of conserved membrane proteins. Here, we present genetic evidence that only one of these two families is essential in Bacillus subtilis .
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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 | 0.000 |
| 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".