The Bardet-Biedl syndrome-related protein CCDC28B modulates mTORC2 function and interacts with SIN1 to control cilia length independently of the mTOR complex

Hum Mol Genet. 2013 Oct 15;22(20):4031-42. doi: 10.1093/hmg/ddt253. Epub 2013 May 31.

Abstract

CCDC28B encodes a coiled coil domain-containing protein involved in ciliogenesis that was originally identified as a second site modifier of the ciliopathy Bardet-Biedl syndrome. We have previously shown that the depletion of CCDC28B leads to shortened cilia; however, the mechanism underlying how this protein controls ciliary length is unknown. Here, we show that CCDC28B interacts with SIN1, a component of the mTOR complex 2 (mTORC2), and that this interaction is important both in the context of mTOR signaling and in a hitherto unknown, mTORC-independent role of SIN1 in cilia biology. We show that CCDC28B is a positive regulator of mTORC2, participating in its assembly/stability and modulating its activity, while not affecting mTORC1 function. Further, we show that Ccdc28b regulates cilia length in vivo, at least in part, through its interaction with Sin1. Importantly, depletion of Rictor, another core component of mTORC2, does not result in shortened cilia. Taken together, our findings implicate CCDC28B in the regulation of mTORC2, and uncover a novel function of SIN1 regulating cilia length that is likely independent of mTOR signaling.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Signal Transducing / metabolism*
  • Animals
  • Bardet-Biedl Syndrome / metabolism*
  • Carrier Proteins / metabolism*
  • Cell Cycle Proteins / metabolism*
  • Cilia / metabolism*
  • Cytoskeletal Proteins
  • Gene Expression Regulation
  • Gene Knockdown Techniques
  • HEK293 Cells
  • HeLa Cells
  • Humans
  • Mechanistic Target of Rapamycin Complex 2
  • Mice
  • Microtubule-Associated Proteins
  • Multiprotein Complexes / metabolism*
  • NIH 3T3 Cells
  • Rapamycin-Insensitive Companion of mTOR Protein
  • Signal Transduction / physiology
  • TOR Serine-Threonine Kinases / metabolism*
  • Zebrafish / embryology
  • Zebrafish / metabolism
  • Zebrafish Proteins / metabolism

Substances

  • Adaptor Proteins, Signal Transducing
  • CCDC28B protein, human
  • CCDC28B protein, mouse
  • Carrier Proteins
  • Cell Cycle Proteins
  • Cytoskeletal Proteins
  • MAPKAP1 protein, human
  • Microtubule-Associated Proteins
  • Multiprotein Complexes
  • RICTOR protein, human
  • Rapamycin-Insensitive Companion of mTOR Protein
  • Zebrafish Proteins
  • rictor protein, mouse
  • stress-activated protein kinase-interacting protein, mouse
  • Mechanistic Target of Rapamycin Complex 2
  • TOR Serine-Threonine Kinases