Congenital Heart Disease Genetics Uncovers Context-Dependent Organization and Function of Nucleoporins at Cilia.
Human genomics is identifying candidate genes for congenital heart disease (CHD), but discovering the underlying mechanisms remains challenging. In a patient with CHD and heterotaxy (Htx), a disorder of left-right patterning, we previously identified a duplication in Nup188. However, a mechanism to explain how a component of the nuclear pore complex (NPC) could cause Htx/CHD was undefined. Here, we show that knockdown of Nup188 or its binding partner Nup93 leads to a loss of cilia during embryonic development while leaving NPC function largely intact. Many data, including the localization of endogenous Nup188/93 at cilia bases, support their direct role at cilia. Super-resolution imaging of Nup188 shows two barrel-like structures with dimensions and organization incompatible with an NPC-like ring, arguing against a proposed "ciliary pore complex." We suggest that the nanoscale organization and function of nucleoporins are context dependent in a way that is required for the structure of the heart.
Program in Vertebrate Developmental Biology, Departments of Pediatrics and Genetics, Yale University School of Medicine, 333 Cedar Street, New Haven, CT 06520, USA.
Florencia
F
Del Viso
Department of Cell Biology, Yale University School of Medicine, 333 Cedar Street, New Haven, CT 06520, USA; Weldon School of Biomedical Engineering, College of Engineering, Purdue University, West Lafayette, IN 47907, USA.
Fang
F
Huang
Department of Cell Biology, Yale University School of Medicine, 333 Cedar Street, New Haven, CT 06520, USA.
Jordan
J
Myers
Department of Cell Biology, Yale University School of Medicine, 333 Cedar Street, New Haven, CT 06520, USA.
Madeleine
M
Chalfant
Department of Cell Biology, Yale University School of Medicine, 333 Cedar Street, New Haven, CT 06520, USA.
Yongdeng
Y
Zhang
Program in Vertebrate Developmental Biology, Departments of Pediatrics and Genetics, Yale University School of Medicine, 333 Cedar Street, New Haven, CT 06520, USA.
Nooreen
N
Reza
Department of Cell Biology, Yale University School of Medicine, 333 Cedar Street, New Haven, CT 06520, USA.
Joerg
J
Bewersdorf
Department of Cell Biology, Yale University School of Medicine, 333 Cedar Street, New Haven, CT 06520, USA. Electronic address: patrick.lusk@yale.edu.
C Patrick
CP
Lusk
Program in Vertebrate Developmental Biology, Departments of Pediatrics and Genetics, Yale University School of Medicine, 333 Cedar Street, New Haven, CT 06520, USA. Electronic address: mustafa.khokha@yale.edu.
Mustafa K
MK
Khokha
D000818:Animals; D002923:Cilia / Q000235:genetics / Q000473:pathology; D055785:Gene Knockdown Techniques; D015894:Genome, Human; D006330:Heart Defects, Congenital / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D059446:Heterotaxy Syndrome / Q000235:genetics* / Q000473:pathology; D006801:Humans; D022022:Nuclear Pore / Q000235:genetics / Q000378:metabolism; D028861:Nuclear Pore Complex Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011487:Protein Conformation; D060066:Protein Interaction Maps / Q000235:genetics; D021381:Protein Transport / Q000235:genetics; D014981:Xenopus / Q000235:genetics; D029867:Xenopus Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
D028861:Nuclear Pore Complex Proteins; C416234:Nup188 protein, Xenopus; C508154:Nup93 protein, human; D029867:Xenopus Proteins
3D nanoscopy; Xenopus tropicalis; congenital heart disease; heterotaxy; left-right patterning; nuclear pore complex; nucleoporin; super-resolution