Identification of regulatory modules mediating specific expression of the roughest gene in Drosophila melanogaster

Abstract

Roughest (Rst) is a cell adhesion molecule of the immunoglobulin superfamily with pleiotropic functions during the development of Drosophila melanogaster. It has been shown to be involved in cell sorting before apoptosis in the developing compound eye, in fusion processes of embryonic muscle development and in axonal pathfinding. In accordance with its multiple functions, the rst gene shows a dynamic expression pattern throughout the development of Drosophila. In order to understand the transcriptional regulation of rst expression we have identified rst cis regulatory sequences in an enhancer detection screen. By dissection of the identified rst cis regulatory sequences we identified several distinct rst regulatory modules. Among others these include elements for expression in interommatidial cells of the pupal eye disc at a time when apoptotic decisions are made in these cells and elements for expression in the embryonic mesoderm. The expression of rst in the embryonic mesoderm is regulated by at least two separate modules.

rst Transcriptional Activity Influences kirre mRNA Concentration in the Drosophila Pupal Retina during the Final Steps of Ommatidial Patterning

“…This delay correlates with late cell sorting, late and erratic apoptotic elimination of surplus IOCs and incomplete pc2 and pc3 differentiation (32). Identification of a genomic rearrangement associated to the rst D allele and its mapping to the rst regulatory region [28], [32] raised the question of how such a mutation could directly affect Rst protein redistribution in IOCs. A relatively simple explanation, based on a combination of differential stability of Rst at the pc1/IOC borders and tightly regulated changes in rst transcription around cell sorting stage was proposed [32].…”

MiMIC: a highly versatile transposon insertion resource for engineering Drosophila melanogaster genes

“…A mutagenic GAL4 gene-trap cassette, encompassing the GAL4 coding sequence and Hsp70 polyadenylation signal, was obtained from plasmid pChs-GAL4 (Drosophila Genomics Resource Center) 55 , and PCR amplified with primers GAL4-Hsp70-EcoRI-F and GAL4-Hsp70-BamHI-R. A mutagenic QF gene-trap cassette, encompassing the QF coding sequence and Hsp70 polyadenylation signal, was obtained from plasmid pattB-QF-Hsp70 (Addgene) 30 , and PCR amplified with primers QF-SV40-EcoRI-F and QF-SV40-BamHI-R. A mutagenic Flp fate mapping gene-trap cassette, encompassing the FLPo 56 coding sequence and SV40 polyadenylation signal, was obtained from plasmid pQUAS-DSCP-Flpo (Addgene) 30 , and PCR amplified with primers Flpo-SV40-EcoRI-F and Flpo-SV40-BamHI-R. The resulting PCR fragments were cut with EcoRI and BamHI and subcloned into pBS-KS-attB1-2-GT-SA , cut with EcoRI and BamHI, resulting in the plasmids pBS-KS-attB1-2-GT-SA-GAL4-Hsp70, pBS-KS-attB1-2-GT-SA-Flp-SV40, and pBS-KS-attB1-2-GT-SA-QF-Hsp70 respectively.…”

Notch Controls Cell Adhesion in the Drosophila Eye

“…rst-Gal4 was obtained from National Institute of Genetics Fly Stock Center (Japan). Other flies used: rst F6 -lacZ [31], spa-Gal4 [25], UAS-N-cadherin [12], sns ZF1.4 and UAS-sns (gift of Susan Abmayr), UAS-N ICD (gift of Cedric Wesley), UAS-N ICD -lexA (gift of Toby Lieber), P[w+]36.1 and hbs 459 (gift of Mary Baylies), UAS-hbs (gift of Helen Sink), GBE-Su(H) m8 -lacZ ( N-lacZ ) [22], Gal-54 [23], UAS-rst (gift of Karl-F. Fischbach), UAS-kirre/duf (gift of Marc Ruiz-Gomez), UAS-Dl (gift of Marek Mlodzik) and hsFLP MKRS (gift of Matthew Freeman).…”