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Wiggan, O.

Publications and source records attributed to Wiggan, O..

2 recordsLinked to original sources

Cell Morphology accurately predicts the nuclear shape of adherent cells

Cells are internally tensed, or prestressed, largely by actomyosin contractility. We hypothesized that nuclear shape is quantitatively predictable from cell shape since prestress couples them both. We trained machine learning models on a publicly available image database of the WTC-11 cell line and predicted shape modes of the nucleus with high accuracy. We develop a U-Net architecture-based model, Cell2Nuc, that predicted nuclear voxels from the cell membrane with accuracies between 74%-87%. To investigate prestress, we cultured and imaged HeLa cells after inhibiting actomyosin contractility. The Cell2Nuc model retrained on the HeLa cells predicted nuclear voxels with slightly lower accuracy. Statistical analysis revealed changes in nuclear size and chromatin organization upon prestress inhibition. Similar trends were seen in images taken from NIH3T3 cells. Thus, cell shape encodes features of nuclear shape, their coupling is partly due to actomyosin contractility, whose abrogation leads to changes in chromatin organization of mechanosensitive origin.

cell biology↗

Single molecule imaging of the central dogma reveals myosin-2A gene expression is regulated by contextual translational buffering

While protein homeostasis is a hallmark of gene regulation, unraveling the hidden regulatory mechanisms that maintain homeostasis is difficult using traditional methods. To confront this problem, we CRISPR engineered a human cell line with multiple tags in the endogenous MYH9 gene, which encodes the essential and ubiquitous myosin-2A cytoskeletal motor. Using these cells, we imaged MYH9 transcription, translation, and mature mRNA and protein in distinct colors, enabling a full dissection of the central dogma. Our data show that MYH9 transcription is upregulated in an SRF-dependent manner in response to cytoskeletal cues and that MYH9 translation can either buffer or match the transcriptional response depending on context. Upon knockdown of actin-depolymerizing proteins like cofilin, translation efficiency drops by a factor of two to buffer strong transcriptional upregulation, likely to help prevent excessive myosin activity. In contrast, following serum stimulation, translation matches the transcriptional response to readily reestablish steady state. Our results identify contextual translational buffering as an important regulatory mechanism driving stable MYH9 expression. They also demonstrate the power and broad applicability of our cell line, which can now be used to accurately quantify central dogma dynamics in response to diverse forms of cellular perturbations.

cell biology↗