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2016 ; 27
(8
): 1397-407
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A genetic interaction map of cell cycle regulators
#MMPMID26912791
Billmann M
; Horn T
; Fischer B
; Sandmann T
; Huber W
; Boutros M
Mol Biol Cell
2016[Apr]; 27
(8
): 1397-407
PMID26912791
show ga
Cell-based RNA interference (RNAi) is a powerful approach to screen for
modulators of many cellular processes. However, resulting candidate gene lists
from cell-based assays comprise diverse effectors, both direct and indirect, and
further dissecting their functions can be challenging. Here we screened a
genome-wide RNAi library for modulators of mitosis and cytokinesis inDrosophilaS2
cells. The screen identified many previously known genes as well as modulators
that have previously not been connected to cell cycle control. We then
characterized ?300 candidate modifiers further by genetic interaction analysis
using double RNAi and a multiparametric, imaging-based assay. We found that
analyzing cell cycle-relevant phenotypes increased the sensitivity for
associating novel gene function. Genetic interaction maps based on mitotic index
and nuclear size grouped candidates into known regulatory complexes of mitosis or
cytokinesis, respectively, and predicted previously uncharacterized components of
known processes. For example, we confirmed a role for theDrosophilaCCR4 mRNA
processing complex componentl(2)NC136during the mitotic exit. Our results show
that the combination of genome-scale RNAi screening and genetic interaction
analysis using process-directed phenotypes provides a powerful two-step approach
to assigning components to specific pathways and complexes.
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