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A Second-Site Noncomplementation Screen for Modifiers of Rho1 Signaling during Imaginal Disc Morpogenesis in Drosophila
Patch, Kistie Stewart, Shannon Welch, Aaron Ward, Robert E., IV
Patch, Kistie
Stewart, Shannon
Welch, Aaron
Ward, Robert E., IV
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Abstract
Background: Rho1 is a small GTPase of the Ras superfamily that serves as the central component in a highly conserved
signaling pathway that regulates tissue morphogenesis during development in all animals. Since there is tremendous
diversity in the upstream signals that can activate Rho1 as well as the effector molecules that carry out its functions, it is
important to define relevant Rho1-interacting genes for each morphogenetic event regulated by this signaling pathway.
Previous work from our lab and others has shown that Rho signaling is necessary for the morphogenesis of leg imaginal
discs during metamorphosis in Drosophila, although a comprehensive identification of Rho1-interacting genes has not been
attempted for this process.
Methodology/Principal Findings: We characterized an amorphic allele of Rho1 that displays a poorly penetrant dominant
malformed leg phenotype and is capable of being strongly enhanced by Rho1-interacting heterozygous mutations. We then
used this allele in a second-site noncomplementation screen with the Exelixis collection of molecularly defined deficiencies
to identify Rho1-interacting genes necessary for leg morphogenesis. In a primary screen of 461 deficiencies collectively
uncovering ,50% of the Drosophila genome, we identified twelve intervals harboring Rho1-interacting genes. Through
secondary screening we identified six Rho1-interacting genes including three that were previously identified (RhoGEF2,
broad, and stubbloid), thereby validating the screen. In addition, we identified Cdc42, Rheb and Sc2 as novel Rho1-interacting
genes involved in adult leg development.
Conclusions/Significance: This screen identified well-known and novel Rho1-interacting genes necessary for leg
morphogenesis, thereby increasing our knowledge of this important signaling pathway. We additionally found that Rheb
may have a unique function in leg morphogenesis that is independent of its regulation of Tor.
Description
Date
2009-10-23
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Public Library of Science
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Keywords
Rho signaling, Drosophila, Morphogenesis, Imaginal disc, Cell shape change
Citation
Patch, K., Stewart, S., Welch, A. and Ward, R. E. 2009. A Second-site Noncomplementation Screen for Modifiers of Rho1 Signaling during Imaginal Disc Morphogenesis in Drosophila. PLoS ONE 4:e7574. http://dx.doi.org/10.1371/journal.pone.0007574