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First published online 4 April 2007
doi: 10.1242/dev.02842

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The F-actin-microtubule crosslinker Shot is a platform for Krasavietz-mediated translational regulation of midline axon repulsion

¹ Department of Cell and Developmental Biology, School of Dentistry, Seoul National University, Seoul 110-740, Republic of Korea.
² School of Biological Sciences, College of Natural Sciences, Seoul National University, Seoul 151-742, Republic of Korea.
³ Department of Cell and Developmental Biology, Vanderbilt University Medical Center, Nashville, TN 37232-2175, USA.
⁴ Department of Physiology, School of Dentistry, Seoul National University, Seoul 110-740, Republic of Korea.

^* Author for correspondence (e-mail: seunglee{at}snu.ac.kr)

Accepted 22 February 2007

Axon extension and guidance require a coordinated assembly of F-actin and microtubules as well as regulated translation. The molecular basis of how the translation of mRNAs encoding guidance proteins could be closely tied to the pace of cytoskeletal assembly is poorly understood. Previous studies have shown that the F-actin-microtubule crosslinker Short stop (Shot) is required for motor and sensory axon extension in the Drosophila embryo. Here, we provide biochemical and genetic evidence that Shot functions with a novel translation inhibitor, Krasavietz (Kra, Exba), to steer longitudinally directed CNS axons away from the midline. Kra binds directly to the C-terminus of Shot, and this interaction is required for the activity of Shot to support midline axon repulsion. shot and kra mutations lead to weak robo-like phenotypes, and synergistically affect midline avoidance of CNS axons. We also show that shot and kra dominantly enhance the frequency of midline crossovers in embryos heterozygous for slit or robo, and that in kra mutant embryos, some Robo-positive axons ectopically cross the midline that normally expresses the repellent Slit. Finally, we demonstrate that Kra also interacts with the translation initiation factor eIF2ß and inhibits translation in vitro. Together, these data suggest that Kra-mediated translational regulation plays important roles in midline axon repulsion and that Shot functions as a direct physical link between translational regulation and cytoskeleton reorganization.

Key words: kra, shot, Translation, Axon guidance, CNS, Drosophila

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