MicroRNA transgene overexpression complements deficiency-based modifier screens in Drosophila

TY - JOUR

T1 - MicroRNA transgene overexpression complements deficiency-based modifier screens in Drosophila

AU - Szuplewski, Sébastien

AU - Kugler, Jan-Michael

AU - Lim, Sing Fee

AU - Verma, Pushpa

AU - Chen, Ya-Wen

AU - Cohen, Stephen M

PY - 2012/2

Y1 - 2012/2

N2 - Dosage-sensitive modifier screening is a powerful tool for linking genes to biological processes. Use of chromosomal deletions permits sampling the effects of removing groups of genes related by position on the chromosome. Here, we explore the use of inducible microRNA transgenes as a complement to deficiency-based modifier screens. miRNAs are predicted to have hundreds of targets. miRNA overexpression provides an efficient means to reduces expression of large gene sets. A collection of transgenes was prepared to allow overexpression of 89 miRNAs or miRNA clusters. These transgenes and a set of genomic deficiencies were screened for their ability to modify the bristle phenotype of the cell-cycle regulator minus. Sixteen miRNAs were identified as dominant suppressors, while the deficiency screen uncovered four genomic regions that contain a dominant suppressor. Comparing the genes uncovered by the deletions with predicted miRNA targets uncovered a small set of candidate suppressors. Two candidates were identified as suppressors of the minus phenotype, Cullin-4 and CG5199/Cut8. Additionally, we show that Cullin-4 acts through its substrate receptor Cdt2 to suppress the minus phenotype. We suggest that inducible microRNA transgenes are a useful complement to deficiency-based modifier screens.

AB - Dosage-sensitive modifier screening is a powerful tool for linking genes to biological processes. Use of chromosomal deletions permits sampling the effects of removing groups of genes related by position on the chromosome. Here, we explore the use of inducible microRNA transgenes as a complement to deficiency-based modifier screens. miRNAs are predicted to have hundreds of targets. miRNA overexpression provides an efficient means to reduces expression of large gene sets. A collection of transgenes was prepared to allow overexpression of 89 miRNAs or miRNA clusters. These transgenes and a set of genomic deficiencies were screened for their ability to modify the bristle phenotype of the cell-cycle regulator minus. Sixteen miRNAs were identified as dominant suppressors, while the deficiency screen uncovered four genomic regions that contain a dominant suppressor. Comparing the genes uncovered by the deletions with predicted miRNA targets uncovered a small set of candidate suppressors. Two candidates were identified as suppressors of the minus phenotype, Cullin-4 and CG5199/Cut8. Additionally, we show that Cullin-4 acts through its substrate receptor Cdt2 to suppress the minus phenotype. We suggest that inducible microRNA transgenes are a useful complement to deficiency-based modifier screens.

KW - Animals

KW - Base Sequence

KW - Caenorhabditis elegans Proteins

KW - Cell Cycle Proteins

KW - Drosophila

KW - Drosophila Proteins

KW - Female

KW - Gene Dosage

KW - Gene Expression

KW - Gene Order

KW - Genetic Vectors

KW - Heat-Shock Proteins

KW - Ligases

KW - Male

KW - MicroRNAs

KW - Molecular Sequence Data

KW - Mutation

KW - Phenotype

KW - Transgenes

U2 - 10.1534/genetics.111.136689

DO - 10.1534/genetics.111.136689

M3 - Journal article

C2 - 22095085

SN - 1943-2631

VL - 190

SP - 617

EP - 626

JO - Genetics

JF - Genetics

IS - 2

ER -