Ift43 Controls the Ciliary Levels of Gli2 and Gli3

Abstract Intraflagellar transport (IFT) drives the bidirectional movement of trains composed of IFT-A, IFT-B, and BBSome complexes that build and maintain cilia while supporting their signaling functions. Over evolution, IFT became integral to Hedgehog signaling by directing the dynamic movements of receptors and Gli transcription factors that fine-tune pathway output. The IFT-A complex contains six subunits, but the smallest, Ift43, remains poorly characterized and is absent from many ciliated species, suggesting specialized roles in signaling rather than core ciliogenesis. Here we show that loss of Ift43 in mice causes mid-gestation lethality with severe craniofacial defects, exencephaly, abdominal wall defects with exposed viscera, edema, and limb patterning defects. At the cellular level, Ift43 deficiency reduces both the number and length of cilia and blocks induction of Gli1 following pathway activation by the agonist SAG. Although Smoothened relocalizes to cilia normally, Ift43 mutants abnormally accumulate Gli2 and Gli3 at ciliary tips before stimulation and continue to generate repressor forms after activation. Conversely, Ift43 overexpression increases basal Gli2 cleavage, revealing an unanticipated role for Ift43 in regulating Gli processing. Together, these findings identify Ift43 as a key IFT-A component that links ciliary assembly to Hedgehog signal transduction and helps set the balance between Gli activator and repressor forms. Competing Interest Statement The authors have declared no competing interest. Footnotes Michael W. Stuck Michael.Stuck{at}umassmed.edu, Mohona Gupta Mohona.Gupta{at}umassmed.edu, Luke N. Knutson luke.knutson{at}jacks.sdstate.edu, Paurav B. Desai Puarav.Desai{at}umassmed.edu, Karyn L. Robert Karyn.Robert1{at}umassmed.edu, Jeffrey J. Anuszczyk Jeffrey.Anuszczyk{at}umassmed.edu, Abigail O. Smith Abigail.Smith{at}umassmed.edu, Delayna Paulson StellpflugPaulson.Delayna{at}mayo.edu, Sumeda Nandadasa Sumeda.Nandadsada{at}umassmed.edu, Hemant Khanna Hemant.Khanna{at}ivericbio.com, William Devine Bill.Devine{at}chp.edu, Cecilia W. Lo Cel36{at}pitt.edu, Timothy Coxcoxtc{at}umkc.edu, Darci M. Fink Darci.Fink{at}sdstate.edu, Gregory J. Pazour Gregory.Pazour{at}umassmed.edu Abbreviations - AVSD - atrioventricular septal defect - BSA - bovine serum albumin - CT - computed tomography - DAPI - 4′,6-diamidino-2-phenylindole - DEXV - dextroversion - EFIC - episcopic fluorescence image capture - E - embryonic day - ERG - electroretinograms - G - Golgi complex - IS - inner segment - IFT - intraflagellar transport - INL - inner nuclear layer - LA - left atria - LV - left ventricle - mVSD - muscular ventricular septal defect - OS - outer segment - ONL - outer nuclear layer - OPL - outer plexiform layer - P - postnatal day - PTA - persistent truncus arteriosus - RA - right atria - ROI - region of interest - RV - right ventricle - SIf - superior-inferior ventricles - TEM - transmission electron microscopy - WGA - Wheat germ agglutinin