NINL and DZANK1 Co-function in Vesicle Transport and Are Essential for Photoreceptor Development in Zebrafish.
Ciliopathies are Mendelian disorders caused by dysfunction of cilia, ubiquitous organelles involved in fluid propulsion (motile cilia) or signal transduction (primary cilia). Retinal dystrophy is a common phenotypic characteristic of ciliopathies since photoreceptor outer segments are specialized primary cilia. These ciliary structures heavily rely on intracellular minus-end directed transport of cargo, mediated at least in part by the cytoplasmic dynein 1 motor complex, for their formation, maintenance and function. Ninein-like protein (NINL) is known to associate with this motor complex and is an important interaction partner of the ciliopathy-associated proteins lebercilin, USH2A and CC2D2A. Here, we scrutinize the function of NINL with combined proteomic and zebrafish in vivo approaches. We identify Double Zinc Ribbon and Ankyrin Repeat domains 1 (DZANK1) as a novel interaction partner of NINL and show that loss of Ninl, Dzank1 or both synergistically leads to dysmorphic photoreceptor outer segments, accumulation of trans-Golgi-derived vesicles and mislocalization of Rhodopsin and Ush2a in zebrafish. In addition, retrograde melanosome transport is severely impaired in zebrafish lacking Ninl or Dzank1. We further demonstrate that NINL and DZANK1 are essential for intracellular dynein-based transport by associating with complementary subunits of the cytoplasmic dynein 1 motor complex, thus shedding light on the structure and stoichiometry of this important motor complex. Altogether, our results support a model in which the NINL-DZANK1 protein module is involved in the proper assembly and folding of the cytoplasmic dynein 1 motor complex in photoreceptor cells, a process essential for outer segment formation and function.
Department of Otorhinolaryngology, Radboud University Medical Centre, Nijmegen, the Netherlands; Radboud Institute for Molecular Life Sciences, Radboud University Nijmegen, Nijmegen, the Netherlands.
Margo
M
Dona
Institute of Molecular Life Sciences, University of Zurich, Zürich, Switzerland; Institute of Medical Genetics, University of Zurich, Zürich, Switzerland.
Ruxandra
R
Bachmann-Gagescu
Division of Experimental Ophthalmology, and Medical Proteome Center, Centre for Ophthalmology, Eberhard Karls University Tuebingen, Tübingen, Germany.
Yves
Y
Texier
Structural and Computational Biology Unit, European Molecular Biology Laboratory, Heidelberg, Germany.
Grischa
G
Toedt
Radboud Institute for Molecular Life Sciences, Radboud University Nijmegen, Nijmegen, the Netherlands; Department of Human Genetics, Radboud University Medical Centre, Nijmegen, the Netherlands.
Lisette
L
Hetterschijt
Department of Otorhinolaryngology, Radboud University Medical Centre, Nijmegen, the Netherlands.
Edith L
EL
Tonnaer
Department of Otorhinolaryngology, Radboud University Medical Centre, Nijmegen, the Netherlands.
Theo A
TA
Peters
Radboud Institute for Molecular Life Sciences, Radboud University Nijmegen, Nijmegen, the Netherlands; Department of Human Genetics, Radboud University Medical Centre, Nijmegen, the Netherlands.
Sylvia E C
SE
van Beersum
Department of Human Genetics, Radboud University Medical Centre, Nijmegen, the Netherlands.
Judith G M
JG
Bergboer
Division of Experimental Ophthalmology, and Medical Proteome Center, Centre for Ophthalmology, Eberhard Karls University Tuebingen, Tübingen, Germany.
Nicola
N
Horn
Department of Otorhinolaryngology, Radboud University Medical Centre, Nijmegen, the Netherlands; Radboud Institute for Molecular Life Sciences, Radboud University Nijmegen, Nijmegen, the Netherlands.
Erik
E
de Vrieze
Department of Otorhinolaryngology, Radboud University Medical Centre, Nijmegen, the Netherlands; Radboud Institute for Molecular Life Sciences, Radboud University Nijmegen, Nijmegen, the Netherlands.
Ralph W N
RW
Slijkerman
Radboud Institute for Molecular Life Sciences, Radboud University Nijmegen, Nijmegen, the Netherlands; Department of Human Genetics, Radboud University Medical Centre, Nijmegen, the Netherlands.
Jeroen
J
van Reeuwijk
Department of Organismal Animal Physiology, Institute for Water and Wetland Research, Radboud University Nijmegen, Nijmegen, The Netherlands.
Gert
G
Flik
Department of Ophthalmology, Radboud University Medical Centre, Nijmegen, the Netherlands.
Jan E
JE
Keunen
Division of Experimental Ophthalmology, and Medical Proteome Center, Centre for Ophthalmology, Eberhard Karls University Tuebingen, Tübingen, Germany.
Marius
M
Ueffing
Structural and Computational Biology Unit, European Molecular Biology Laboratory, Heidelberg, Germany.
Toby J
TJ
Gibson
Radboud Institute for Molecular Life Sciences, Radboud University Nijmegen, Nijmegen, the Netherlands; Department of Human Genetics, Radboud University Medical Centre, Nijmegen, the Netherlands.
Ronald
R
Roepman
Division of Experimental Ophthalmology, and Medical Proteome Center, Centre for Ophthalmology, Eberhard Karls University Tuebingen, Tübingen, Germany.
Karsten
K
Boldt
Department of Otorhinolaryngology, Radboud University Medical Centre, Nijmegen, the Netherlands; Radboud Institute for Molecular Life Sciences, Radboud University Nijmegen, Nijmegen, the Netherlands; Department of Human Genetics, Radboud University Medical Centre, Nijmegen, the Netherlands.
Hannie
H
Kremer
Department of Otorhinolaryngology, Radboud University Medical Centre, Nijmegen, the Netherlands; Radboud Institute for Molecular Life Sciences, Radboud University Nijmegen, Nijmegen, the Netherlands.
Erwin
E
van Wijk
D000818:Animals; D001692:Biological Transport / Q000235:genetics; D002352:Carrier Proteins / Q000235:genetics*; D002923:Cilia / Q000235:genetics; D004398:Dyneins / Q000235:genetics*; D057809:HEK293 Cells; D006801:Humans; D007814:Larva / Q000235:genetics* / Q000254:growth & development; D008869:Microtubule-Associated Proteins / Q000235:genetics*; D055495:Neurogenesis / Q000235:genetics; D009687:Nuclear Proteins / Q000235:genetics*; D020419:Photoreceptor Cells, Vertebrate; D040901:Proteomics; D012160:Retina / Q000254:growth & development*; D015398:Signal Transduction; D015027:Zebrafish / Q000235:genetics / Q000254:growth & development; D029961:Zebrafish Proteins / Q000235:genetics*
D002352:Carrier Proteins; C000607992:DZANK1 protein, zebrafish; D008869:Microtubule-Associated Proteins; C476755:NINL protein, human; D009687:Nuclear Proteins; D029961:Zebrafish Proteins; D004398:Dyneins