@article{12511da5847d488daaf671aee8b17c42,
title = "Ankmy2 Prevents Smoothened-Independent Hyperactivation of the Hedgehog Pathway via Cilia-Regulated Adenylyl Cyclase Signaling",
abstract = "The mechanisms underlying subcellular targeting of cAMP-generating adenylyl cyclases and processes regulated by their compartmentalization are poorly understood. Here, we identify Ankmy2 as a repressor of the Hedgehog pathway via adenylyl cyclase targeting. Ankmy2 binds to multiple adenylyl cyclases, determining their maturation and trafficking to primary cilia. Mice lacking Ankmy2 are mid-embryonic lethal. Knockout embryos have increased Hedgehog signaling and completely open neural tubes showing co-expansion of all ventral neuroprogenitor markers, comparable to the loss of the Hedgehog receptor Patched1. Ventralization in Ankmy2 knockout is completely independent of the Hedgehog pathway transducer Smoothened. Instead, ventralization results from the reduced formation of Gli2 and Gli3 repressors and early depletion of adenylyl cyclase III in neuroepithelial cilia, implicating deficient pathway repression. Ventralization in Ankmy2 knockout requires both cilia and Gli2 activation. These findings indicate that cilia-dependent adenylyl cyclase signaling represses the Hedgehog pathway and promotes morphogenetic patterning.",
keywords = "ACIII, Ankmy2, Gli repressor, Smoothened, adenylyl cyclase, cAMP, neural tube, primary cilia, protein kinase A, sonic hedgehog",
author = "Somatilaka, {Bandarigoda Nipunika} and Hwang, {Sun Hee} and Palicharla, {Vivek Reddy} and White, {Kevin Andrew} and Hemant Badgandi and Shelton, {John Michael} and Saikat Mukhopadhyay",
note = "Funding Information: This project was funded by Alex's Lemonade Foundation (A-grant to S.M.), Welch Foundation (grant #I-1906 to S.M.), and National Institutes of Health (1R01GM113023 to S.M. and S10RR029731 to Live Cell Imaging Facility, UT Southwestern). We thank UT Southwestern's transgenic, molecular pathology, electron microscopy and mass spectrometry cores, and mouse animal care facility. We thank Ross Tomaino (Taplin Mass Spectrometry Facility) and Andrew Lemoff (UT Southwestern proteomics core) for assistance with mass spectrometry and Cameron Perry for paraffin sectioning. We acknowledge gifts of reagents from Tamara Caspary, Kathryn Anderson, Jonathan Eggenschwiler, Ron Taussig, Andrew McMahon, Deanna Grant, Andrew Peterson, Thomas Carroll, Peter Michaely, Rhonda Bassel-Duby, and Eric Olson. We thank Sandra Schmid and Sandii Constable for comments on the manuscript and Karel Liem for advice on immunostaining of ACs. Monoclonal antibodies developed by T.M. Jessell/S. Brenner-Morton (Nkx2.2), O.D. Madsen (Nkx6.1), and A. Kawakami (Pax7) were obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the NICHD and maintained by the Department of Biological Sciences, the University of Iowa, Iowa City, IA 52242, USA. B.N.S. and S.M. conceived the project, designed experiments, analyzed most of the data, and wrote the paper with inputs from all authors. S.-H.H. developed the mouse model, and B.N.S. and S.-H.H. performed most of the experiments. V.R.P. and K.A.W. performed tandem immunopurification experiments. H.B. and V.R.P. generated mass spectrometry data. J.M.S. performed radiometric in situ hybridization. The authors declare no competing interests. Funding Information: This project was funded by Alex{\textquoteright}s Lemonade Foundation (A-grant to S.M.), Welch Foundation (grant # I-1906 to S.M.), and National Institutes of Health ( 1R01GM113023 to S.M. and S10RR029731 to Live Cell Imaging Facility, UT Southwestern). We thank UT Southwestern{\textquoteright}s transgenic, molecular pathology, electron microscopy and mass spectrometry cores, and mouse animal care facility. We thank Ross Tomaino (Taplin Mass Spectrometry Facility) and Andrew Lemoff (UT Southwestern proteomics core) for assistance with mass spectrometry and Cameron Perry for paraffin sectioning. We acknowledge gifts of reagents from Tamara Caspary, Kathryn Anderson, Jonathan Eggenschwiler, Ron Taussig, Andrew McMahon, Deanna Grant, Andrew Peterson, Thomas Carroll, Peter Michaely, Rhonda Bassel-Duby, and Eric Olson. We thank Sandra Schmid and Sandii Constable for comments on the manuscript and Karel Liem for advice on immunostaining of ACs. Monoclonal antibodies developed by T.M. Jessell/S. Brenner-Morton (Nkx2.2), O.D. Madsen (Nkx6.1), and A. Kawakami (Pax7) were obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the NICHD and maintained by the Department of Biological Sciences, the University of Iowa, Iowa City, IA 52242, USA. Publisher Copyright: {\textcopyright} 2020 Elsevier Inc.",
year = "2020",
month = sep,
day = "28",
doi = "10.1016/j.devcel.2020.06.034",
language = "English (US)",
volume = "54",
pages = "710--726.e8",
journal = "Developmental cell",
issn = "1534-5807",
publisher = "Cell Press",
number = "6",
}