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Published on: April 14, 2023
A Drosophila overexpression screen for modifiers of Rho signalling in cytokinesis
Stephen L Gregory1, Tetyana Shandala, Louise O'Keefe
1ARC Special Research Centre for the Molecular Genetics of Development, University of Adelaide, Adelaide, Australia.
Abstract:
To identify genes that modulate Rho signalling during cytokinesis we tested the effect of overexpressing a set of 2190 genes on an eye phenotype caused by defective Rho activation. The resulting 112 modifier loci fell into three main classes: cell cycle genes, signalling effectors and metabolic enzymes. We developed a further series of genetic tests to refine the interactors into those most likely to modify Rho signalling during cytokinesis. In addition to a number of genes previously implicated in the Rho pathway during cytokinesis, we identified four novel primary candidates: cdc14, Pitslre, PDK1 and thread/diap1. cdc14 orthologs have, however, been implicated in cytokinesis in other organisms, as have molecules related to Thread/Diap1. The identification of several modifiers that are genetically redundant paralogs highlights the ability of overexpression screens to identify genes that are refractory to traditional loss-of-function approaches. Overexpression screens and sensitized phenotypes, therefore, may help identify the many factors that are expected to be involved in cytokinesis but have not been discovered by previous genetic screens.
Insights
This study identified genes controlling cell division (cytokinesis) by overexpressing genes and observing effects on a Rho signaling defect. Four novel candidate genes were found, highlighting overexpression screens for discovering essential cell division factors.
Area of Science:
- Cell Biology
- Genetics
- Molecular Biology
Background:
- Rho signaling is crucial for cytokinesis, the process of cell division.
- Identifying genes that regulate Rho signaling during cytokinesis is essential for understanding cell cycle control.
Purpose of the Study:
- To identify novel genes that modulate Rho signaling during cytokinesis.
- To refine interactors and pinpoint primary candidates involved in Rho pathway regulation.
Main Methods:
- Overexpression screen of 2190 genes using a sensitized eye phenotype caused by defective Rho activation.
- Genetic tests to refine identified modifier loci.
- Analysis of gene classes including cell cycle, signaling, and metabolic enzymes.
Main Results:
- 112 modifier loci were identified, categorized into cell cycle genes, signaling effectors, and metabolic enzymes.
- Four novel primary candidate genes were identified: cdc14, Pitslre, PDK1, and thread/diap1.
- Overexpression screens successfully identified genes refractory to loss-of-function approaches, including genetically redundant paralogs.
Conclusions:
- Overexpression screens are valuable for discovering genes involved in cytokinesis, especially those missed by traditional methods.
- The identified novel candidates (cdc14, Pitslre, PDK1, thread/diap1) offer new insights into Rho signaling during cell division.
- This research expands the known genetic network regulating cytokinesis.
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