Abstract
Bone morphogenetic proteins (BMPs), a subgroup of the transforming growth factor (TGF)-β family, transduce their signal through multiple components downstream of their receptors. Even though the components involved in the BMP signaling pathway have been intensely studied, many molecules mediating BMP signaling remain to be addressed. To identify novel components that participate in BMP signaling, RNA interference (RNAi)-based screening was established by detecting phosphorylated Mad (pMad) in Drosophila S2 cells. Ter94, a member of the family of AAA ATPases, was identified as a novel mediator of BMP signaling, which is required for the phosphorylation of Mad in Drosophila S2 cells. Moreover, the mammalian orthlog of Ter94 valosin-containing protein (VCP) plays a critical role in the BMP-Smad1/5/8 signaling pathway in mammalian cells. Genetic evidence suggests that Ter94 is involved in the dorsal-ventral patterning of the Drosophila early embryo through regulating decapentaplegic (Dpp)/BMP signals. Taken together, our data suggest that Ter94/VCP appears to be an evolutionarily conserved component that regulates BMP-Smad1/5/8 signaling.
Highlights
Understanding tissue generation during metazoan development requires knowledge of how cell and tissue identity is established
To optimize the conditions for largescale screening, images of cells stained with Flag and phosphorylated Mad (pMad) antibodies in 384 multiwell plates were obtained with the high throughput fluorescent microscope Arrayscan, and the protocol was established for detecting bone morphogenetic proteins (BMPs) signals in S2 cells (Fig. 1C)
We established a high throughput RNA interference (RNAi) screening in Drosophila S2 cells to identify novel components involved in BMP signaling
Summary
Understanding tissue generation during metazoan development requires knowledge of how cell and tissue identity is established. Central to this issue is the characterization of signaling molecules, cell-cell interactions, receptors, and second messenger systems that contribute to the processes of cell differentiation and specification. The transforming growth factor (TGF)-b family represents the PLOS ONE | DOI:10.1371/journal.pone.0114475. Ter94/VCP and BMP Signaling largest collection of growth factors identified to date and consists of more than 30 secreted polypeptides. Within this family, the bone morphogenetic proteins (BMPs) constitute the largest subgroup. BMPs regulate biological processes as diverse as cell proliferation, differentiation, cell-fate determination, apoptosis, and morphogenesis [1, 2]
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
