An overexpression screen in Drosophila for genes that restrict growth or cell-cycle progression in the developing eye.
We screened for genes that, when overexpressed in the proliferating cells of the eye imaginal disc, result in a reduction in the size of the adult eye. After crossing the collection of 2296 EP lines to the ey-GAL4 driver, we identified 46 lines, corresponding to insertions in 32 different loci, that elicited a small eye phenotype. These lines were classified further by testing for an effect in postmitotic cells using the sev-GAL4 driver, by testing for an effect in the wing using en-GAL4, and by testing for the ability of overexpression of cycE to rescue the small eye phenotype. EP lines identified in the screen encompass known regulators of eye development including hh and dpp, known genes that have not been studied previously with respect to eye development, as well as 19 novel ORFs. Lines with insertions near INCENP, elB, and CG11518 were characterized in more detail with respect to changes in growth, cell-cycle phasing, and doubling times that were elicited by overexpression. RNAi-induced phenotypes were also analyzed in SL2 cells. Thus overexpression screens can be combined with RNAi experiments to identify and characterize new regulators of growth and cell proliferation.
Tseng
Ai-Sun Kelly
AS
Massachusetts General Hospital Cancer Center and Harvard Medical School, Charlestown, Massachusetts 02129, USA.
Hariharan
Iswar K
IK
D000818:Animals; D015152:Blotting, Northern; D002453:Cell Cycle / Q000235:genetics*; D004330:Drosophila / Q000201:enzymology / Q000235:genetics*; D005123:Eye / Q000166:cytology / Q000196:embryology*; D020869:Gene Expression Profiling; D007150:Immunohistochemistry; D017403:In Situ Hybridization