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Exploring endocytic compartment morphology with systematic genetics and single cell image analysis

View ORCID ProfileMojca Mattiazzi Usaj, Nil Sahin, Helena Friesen, Carles Pons, Matej Usaj, Myra Paz Masinas, Ermira Shuteriqi, Aleksei Shkurin, Patrick Aloy, Quaid Morris, Charles Boone, Brenda J. Andrews
doi: https://doi.org/10.1101/724989
Mojca Mattiazzi Usaj
1The Donnelly Centre, University of Toronto, Toronto, Canada
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  • ORCID record for Mojca Mattiazzi Usaj
Nil Sahin
1The Donnelly Centre, University of Toronto, Toronto, Canada
2Department of Molecular Genetics, University of Toronto, Toronto, Canada
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Helena Friesen
1The Donnelly Centre, University of Toronto, Toronto, Canada
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Carles Pons
3Institute for Research in Biomedicine (IRB Barcelona), the Barcelona Institute for Science and Technology, Barcelona, Catalonia, Spain
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Matej Usaj
1The Donnelly Centre, University of Toronto, Toronto, Canada
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Myra Paz Masinas
1The Donnelly Centre, University of Toronto, Toronto, Canada
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Ermira Shuteriqi
1The Donnelly Centre, University of Toronto, Toronto, Canada
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Aleksei Shkurin
1The Donnelly Centre, University of Toronto, Toronto, Canada
2Department of Molecular Genetics, University of Toronto, Toronto, Canada
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Patrick Aloy
3Institute for Research in Biomedicine (IRB Barcelona), the Barcelona Institute for Science and Technology, Barcelona, Catalonia, Spain
4Institució Catalana de Recerca i Estudis Avançats (ICREA), Barcelona, Catalonia, Spain
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Quaid Morris
1The Donnelly Centre, University of Toronto, Toronto, Canada
2Department of Molecular Genetics, University of Toronto, Toronto, Canada
5Department of Computer Science, University of Toronto, Toronto, Canada
6Vector Institute, Toronto, Canada
7Ontario Institute of Cancer Research, Toronto, Canada
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  • For correspondence: brenda.andrews@utoronto.ca charlie.boone@utoronto.ca quaid.morris@utoronto.ca
Charles Boone
1The Donnelly Centre, University of Toronto, Toronto, Canada
2Department of Molecular Genetics, University of Toronto, Toronto, Canada
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  • For correspondence: brenda.andrews@utoronto.ca charlie.boone@utoronto.ca quaid.morris@utoronto.ca
Brenda J. Andrews
1The Donnelly Centre, University of Toronto, Toronto, Canada
2Department of Molecular Genetics, University of Toronto, Toronto, Canada
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  • For correspondence: brenda.andrews@utoronto.ca charlie.boone@utoronto.ca quaid.morris@utoronto.ca
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ABSTRACT

Endocytosis is a conserved process that mediates the internalization of nutrients and plasma membrane components, including receptors, for sorting to endosomes and the vacuole (lysosome). We combined systematic yeast genetics, high-content screening, and neural network-based image analysis of single cells to screen for genes that influence the morphology of four main endocytic compartments: coat proteins, actin patches, late endosome, and vacuole. This unbiased approach identified 17 mutant phenotypes and ∼1600 genes whose perturbation affected at least one of the four compartments. Numerous mutants were associated with multiple phenotypes, indicating that morphological pleiotropy is often seen within the endocytic pathway. Morphological profiles based on the 17 aberrant phenotypes were highly correlated for functionally related genes, enabling prediction of gene function. Incomplete penetrance was prevalent, and single-cell analysis enabled exploration of the mechanisms underlying cellular heterogeneity, which include replicative age, organelle inheritance, and stress response.

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Posted August 04, 2019.
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Exploring endocytic compartment morphology with systematic genetics and single cell image analysis
Mojca Mattiazzi Usaj, Nil Sahin, Helena Friesen, Carles Pons, Matej Usaj, Myra Paz Masinas, Ermira Shuteriqi, Aleksei Shkurin, Patrick Aloy, Quaid Morris, Charles Boone, Brenda J. Andrews
bioRxiv 724989; doi: https://doi.org/10.1101/724989
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Exploring endocytic compartment morphology with systematic genetics and single cell image analysis
Mojca Mattiazzi Usaj, Nil Sahin, Helena Friesen, Carles Pons, Matej Usaj, Myra Paz Masinas, Ermira Shuteriqi, Aleksei Shkurin, Patrick Aloy, Quaid Morris, Charles Boone, Brenda J. Andrews
bioRxiv 724989; doi: https://doi.org/10.1101/724989

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