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Oesterle, R.

Publications and source records attributed to Oesterle, R..

2 recordsLinked to original sources

Love-thy-neighbor: Neural networks for tracking and lineage tracing in budding yeast

Tracking and lineage tracing are widely needed tasks in biological image analysis. For cells that grow and divide, tracking is challenging because cells change in number, shape, and size throughout a recording. As the time interval between images increases, it becomes more difficult to establish correspondences between cells across timepoints. Consequently, tracking has to be performed between consecutive or temporally close images, which leads to exponentially decreasing tracking accuracy and thus high sensitivity to error rates. For budding yeast, this challenge is further heightened by the similarity of cells in colonies, their dense packing, the asymmetric nature of cell divisions, and movement due to growth of the colony. A related task, lineage tracing, is similarly challenging without fluorescent markers due to multiple potential mother cells surrounding a new daughter cell. Here, we present neural networks for budding yeast tracking and lineage tracing, named LYN-track and LYN-trace, respectively. These methods leverage fine geometric features of cells and their neighborhoods. To train and test the algorithms, we recorded and annotated new budding and fission yeast microscopy movies (78,852 frame-to-frame tracklets, 2,512 images), which we make freely available. On these and existing datasets, our neural network-based methods demonstrate robust, above state-of-the-art performance. Both tools have been integrated into graphical user interfaces (GUIs), available on Github, and can be straightforwardly retrained with custom data if desired.

bioinformatics↗

Why budding yeast overrides the DNA damage checkpoint

Checkpoints arrest biological processes and enhance chances for error correction. In many species ranging from budding yeast to human, checkpoints are eventually overridden despite persistent dam-age. Whether checkpoint override serves a biological function remains unclear. Here, we investigate this question in the context of the DNA damage checkpoint (DDC) in budding yeast. To demonstrate that DDC override increases fitness, we pursued a novel approach: To avoid inherent ambiguities when comparing genetic mutants, we instead employed a light-controlled trigger to finely tune the timing of checkpoint override in a consistent wild-type DDC and DNA repair background. We show that override is beneficial and wild-type override timing maximizes fitness. We formulate two specific hypotheses to explain the fitness benefit: i) override enables multiple rounds of replication, including of broken chromosomal fragments, statistically increasing the chance of at least one successful repair; or ii) override may enhance specific DNA repair pathways. Testing the first hypothesis, we tracked broken chromosome fragments using an optogenetic reporter and found their segregation pattern to be inconsistent with a probabilistic increase in post-override repair opportunities. To test the second hypothesis, we dynamically depleted key repair pathway proteins - individually and combinatorially - without interfering with the establishment of checkpoint arrest. Strikingly, we found that proteins in-volved in microhomology-mediated end joining (MMEJ) substantially enhanced override-associated break repair. Together, these results provide direct evidence of a fitness advantage conferred by checkpoint override and uncover MMEJ-associated repair proteins as the mechanistic basis.

molecular biology↗