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Costal 2 interactions with Cubitus interruptus (Ci) underlying Hedgehog-regulated Ci processing.

Qianhe Zhou1, Daniel Kalderon

  • 1Department of Biological Sciences, Columbia University, 1212 Amsterdam Ave.,New York, NY 10027, USA.qz2107@columbia.edu

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Hedgehog (Hh) signaling regulates cell behavior by controlling transcription factors. This study reveals Cos2 binding to the CORD domain of Ci-155 relieves inhibition, promoting Hh-regulated proteolysis.

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Area of Science:

  • Molecular Biology
  • Developmental Biology
  • Cell Signaling

Background:

  • Extracellular Hedgehog (Hh) proteins regulate cellular behaviors via Gli/Ci transcription factors.
  • Hh signaling in Drosophila involves regulating the proteolytic processing of Ci-155 to Ci-75.
  • Processing is mediated by Cos2 binding to Ci-155, enabling phosphorylation and ubiquitination.

Purpose of the Study:

  • To investigate the interaction domains of Cos2 and Ci-155.
  • To elucidate the role of the CORD domain in Ci-155 processing.
  • To understand the mechanism of Hh-regulated proteolysis.

Main Methods:

  • In vitro binding assays to identify Cos2 interaction sites on Ci-155.
  • Analysis of Ci-155 processing in Drosophila wing discs using Cos2 variants.
  • Investigating the effect of nucleotides on Cos2-Ci interaction and processing.

Main Results:

  • Cos2 binds to the zinc finger region of Ci-155 in addition to CDN and CORD domains.
  • The zinc finger region, along with CORD, redundantly promotes Hh-regulated Ci-155 proteolysis.
  • Cos2 binding to CORD is nucleotide-dependent and abrogated by Cos2 S182N mutation.
  • Deletion of the CORD region enhances processing, and Cos2 S182N stimulates processing upon CORD deletion.

Conclusions:

  • The CORD region of Ci-155 possesses an inhibitory function on processing, distinct from Cos2 binding.
  • Cos2 binding to CORD relieves this inhibition, suggesting a regulatory role.
  • This regulation likely involves a specific nucleotide-bound conformation of Cos2, potentially modulated by Hh signaling.