LRRC6 Mutation Causes Primary Ciliary Dyskinesia with Dynein Arm Defects

Horani, Amjad; Ferkol, Thomas; Shoseyov, David; Wasserman, Mollie G.; Oren, Yifat S.; Kerem, Batsheva; Amirav, Israel; Cohen‐Cymberknoh, Malena; Dutcher, Susan K.; Brody, Steven L.; Elpeleg, Orly; Kerem, Eitan

doi:10.1371/journal.pone.0059436

Cited by 95 publications

(75 citation statements)

References 52 publications

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“…We further show that Reptin directly interact with Lrrc6/ Seahorse, a protein very recently linked to PCD (18,45). Consistent with this physical interaction, mutants of reptin and lrrc6/seahorse were identified in the same genetic screen in zebrafish and both show similar body curvature and kidney cyst formation (22,23).…”

Section: Discussionsupporting

confidence: 75%

Reptin/Ruvbl2 is a Lrrc6/Seahorse interactor essential for cilia motility

Lei

Yuan

Cao

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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Primary ciliary dyskinesia (PCD) is an autosomal recessive disease caused by defective cilia motility. The identified PCD genes account for about half of PCD incidences and the underlying mechanisms remain poorly understood. We demonstrate that Reptin/ Ruvbl2, a protein known to be involved in epigenetic and transcriptional regulation, is essential for cilia motility in zebrafish. We further show that Reptin directly interacts with the PCD protein Lrrc6/Seahorse and this interaction is critical for the in vivo function of Lrrc6/Seahorse in zebrafish. Moreover, whereas the expression levels of multiple dynein arm components remain unchanged or become elevated, the density of axonemal dynein arms is reduced in reptin hi2394 mutants. Furthermore, Reptin is highly enriched in the cytosol and colocalizes with Lrrc6/Seahorse. Combined, these results suggest that the Reptin-Lrrc6/Seahorse complex is involved in dynein arm formation. We also show that although the DNA damage response is induced in reptin hi2394 mutants, it remains unchanged in cilia biogenesis mutants and lrrc6/seahorse mutants, suggesting that increased DNA damage response is not intrinsic to ciliary defects and that in vertebrate development, Reptin functions in multiple processes, both cilia specific and cilia independent.ciliopathy | chromatin remodeling complex | dynein arm assembly factor A lthough the sensory function of the cilium has garnered significant attention only in the past decade, defective cilia motility was recognized nearly four decades ago as the cause of primary (genetic) ciliary dyskinesia (PCD) (1). PCD is a group of rare human genetic diseases characterized by recurrent infections of the respiratory system, male infertility, and frequently laterality defects, all of which are tightly linked to cilia motility abnormalities. Up to now, 19 PCD genes have been identified (2-20). However, combined they account for about 50% of all PCD cases, suggesting the existence of multiple additional causative genes. Despite such significant advances, our understanding of how cilia motility is regulated is limited and the list of genes involved in cilia motility remains incomplete.In motile cilia or flagella, dynein arms are large protein complexes powering cilia motility. Not surprisingly, multiple PCD genes encode dynein arm components, including DNAL1, DNAI1, DNAI2, DNAH5, and DNAH11 (2-6). In addition, it is understood that dynein arm subunits are preassembled in the cytosol, transported into cilia/flagellum, and docked onto the axoneme, although the underlying mechanisms are poorly understood (21). In recent years, three PCD genes, i.e., DNAAF1/LRRC50 (7, 8), DNAAF2/ KTU (9), and DNAAF3/PF22 (10) have been shown to be involved in the assembly of dynein arm subunits, highlighting the relevance and importance of this process in cilia motility and PCD.Very recently, mutations in human LRRC6/SEAHORSE were linked to PCD (18). Similar to the three known dynein arm assembly factors, mutations in LRRC6/SEAHORSE lead to the absence of both the outer...

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Section: Discussionsupporting

confidence: 75%

Reptin/Ruvbl2 is a Lrrc6/Seahorse interactor essential for cilia motility

Lei

Yuan

Cao

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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“…Next, we obtained a previously described insertional mutant, ift172 hi2211tg (Sun et al, 2004), and performed a complementation test; we confirmed that ccn1.9 pd1085 is a mutated allele of ift172. Similarly,dx33.9 pd1087 mapped to an interval containing lrrc6, which encodes a dynein arm ciliary component (Horani et al, 2013), that bears a T-to-G nonsense…”

Section: Implementation Of the Wes Methods For The Identification Of Mmentioning

confidence: 99%

Rapid identification of kidney cyst mutations by whole exome sequencing in zebrafish

et al. 2013

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SUMMARYForward genetic approaches in zebrafish have provided invaluable information about developmental processes. However, the relative difficulty of mapping and isolating mutations has limited the number of new genetic screens. Recent improvements in the annotation of the zebrafish genome coupled to a reduction in sequencing costs prompted the development of whole genome and RNA sequencing approaches for gene discovery. Here we describe a whole exome sequencing (WES) approach that allows rapid and costeffective identification of mutations. We used our WES methodology to isolate four mutations that cause kidney cysts; we identified novel alleles in two ciliary genes as well as two novel mutants. The WES approach described here does not require specialized infrastructure or training and is therefore widely accessible. This methodology should thus help facilitate genetic screens and expedite the identification of mutants that can inform basic biological processes and the causality of genetic disorders in humans.

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“…35,60,61 Gene-specific shRNA sequences subcloned into the pLKO.1 transfer plasmid 62 were from the RNAi Consortium collection including a control nontargeted sequence (SHC016), 2 ATG5-specific sequences (#1, TRCN0000330392; #2, TRCN0000330394) and one ATG14-specific sequence (TRCN0000010211), all available from Sigma-Aldrich. Recombinant lentiviruses were produced by co-transfection of the specific pLKO.1 plasmid that includes a cassette for puromycin resistance with plasmids containing packaging (pHR'8.2DR) and envelope (pCMV-VSV-G) sequences in HEK293T cells using FuGENE 6 (Promega, E2691) as described.…”

Section: Rna Interferencementioning

confidence: 99%

IL13 activates autophagy to regulate secretion in airway epithelial cells

et al. 2016

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Cytokine modulation of autophagy is increasingly recognized in disease pathogenesis, and current concepts suggest that type 1 cytokines activate autophagy, whereas type 2 cytokines are inhibitory. However, this paradigm derives primarily from studies of immune cells and is poorly characterized in tissue cells, including sentinel epithelial cells that regulate the immune response. In particular, the type 2 cytokine IL13 (interleukin 13) drives the formation of airway goblet cells that secrete excess mucus as a characteristic feature of airway disease, but whether this process is influenced by autophagy was undefined. Here we use a mouse model of airway disease in which IL33 (interleukin 33) stimulation leads to IL13-dependent formation of airway goblet cells as tracked by levels of mucin MUC5AC (mucin 5AC, oligomeric mucus/gel forming), and we show that these cells manifest a block in mucus secretion in autophagy gene Atg16l1-deficient mice compared to wild-type control mice. Similarly, primary-culture human tracheal epithelial cells treated with IL13 to stimulate mucus formation also exhibit a block in MUC5AC secretion in cells depleted of autophagy gene ATG5 (autophagy-related 5) or ATG14 (autophagyrelated 14) compared to nondepleted control cells. Our findings indicate that autophagy is essential for airway mucus secretion in a type 2, IL13-dependent immune disease process and thereby provide a novel therapeutic strategy for attenuating airway obstruction in hypersecretory inflammatory diseases such as asthma, chronic obstructive pulmonary disease, and cystic fibrosis lung disease. Taken together, these observations suggest that the regulation of autophagy by Th2 cytokines is cell-context dependent.

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LRRC6 Mutation Causes Primary Ciliary Dyskinesia with Dynein Arm Defects

Cited by 95 publications

References 52 publications

Reptin/Ruvbl2 is a Lrrc6/Seahorse interactor essential for cilia motility

Reptin/Ruvbl2 is a Lrrc6/Seahorse interactor essential for cilia motility

Rapid identification of kidney cyst mutations by whole exome sequencing in zebrafish

IL13 activates autophagy to regulate secretion in airway epithelial cells

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