Abstract
BackgroundBMP signaling is involved in myriad metazoan developmental processes, and study of this pathway in Drosophila has contributed greatly to our understanding of its molecular and genetic mechanisms. These studies have benefited not only from Drosophila’s advanced genetic tools, but from complimentary in vitro culture systems. However, the commonly-used S2 cell line is not intrinsically sensitive to the major BMP ligand Dpp and must therefore be augmented with exogenous pathway components for most experiments.ResultsHerein we identify and characterize the responses of Drosophila ML-DmD17-c3 cells, which are sensitive to Dpp stimulation and exhibit characteristic regulation of BMP target genes including Dad and brk. Dpp signaling in ML-DmD17-c3 cells is primarily mediated by the receptors Put and Tkv, with additional contributions from Wit and Sax. Furthermore, we report complex regulatory feedback on core pathway genes in this system.ConclusionsNative ML-DmD17-c3 cells exhibit robust transcriptional responses to BMP pathway induction. We propose that ML-DmD17-c3 cells are well-suited for future BMP pathway analyses.
Highlights
Bone Morphogenetic Protein (BMP) signaling is involved in myriad metazoan developmental processes, and study of this pathway in Drosophila has contributed greatly to our understanding of its molecular and genetic mechanisms
We demonstrate the respective contributions of the four BMP receptors to signaling, and examine the intricate transcriptional feedback that results from pathway activation in these cells
Identification of ML-DmD17-c3 cells and characterization of their responsiveness to Dpp stimulation Leveraging the transcriptome datasets produced by the modENCODE project [34, 36], we selected three candidate cells lines (ML-DmD4-c1; ML-DmD8; ML-DmD17-c3; [35]) with the highest transcript levels of key components of the Dpp signal transduction cascade (Fig. 1a, Additional file 1: Table S1)
Summary
BMP signaling is involved in myriad metazoan developmental processes, and study of this pathway in Drosophila has contributed greatly to our understanding of its molecular and genetic mechanisms. These studies have benefited from Drosophila’s advanced genetic tools, but from complimentary in vitro culture systems. The Bone Morphogenetic Protein (BMP) signaling pathway plays key roles in metazoan development and stem cell maintenance, at wound healing sites, and in myriad other contexts [1,2,3]. The activated receptor complex recruits and phosphorylates an intracellular signal transduction component, the receptor-regulated R-SMAD transcription factor Mad (Mothers against DPP) [16]. The simplicity of the cascade and the powers of genetic manipulation in Drosophila render the fruit fly a premier system for the study of fundamental aspects of BMP signaling in vivo
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