A Phylogenetic Profiling Approach Identifies Novel Ciliogenesis Genes InDrosophilaAndC. elegans

Abstract

Cilia are cellular projections that perform sensory and motile functions in eukaryotic cells. A defining feature of cilia is that they are evolutionarily ancient yet not universally conserved. In this study we have used the resulting presence and absence pattern in the genomes of diverse eukaryotes to identify a set of 386 human genes associated with cilium assembly or motility. Comprehensive tissue-specific RNAi in Drosophila and mutant analysis in C. elegans revealed signature ciliary defects for 70-80% of novel genes, a percentage similar to that for known genes within the cluster. Further characterization identified different phenotypic classes, including a set of genes related to the cartwheel component Bld10/Cep135 and two highly conserved regulators of cilium biogenesis. We believe this dataset to define the core set of genes required for cilium assembly and motility across eukaryotes, an invaluable resource for future studies of cilium biology and associated disorders.

Centriolar satellites are sites of translation of centrosomal and ciliary proteins

“…Both types of defect could be reproduced by tissue-specific RNAi and rescued by introduction of a GFP transgene, confirming specificity of the mutant phenotype and functionality of the GFP transgene. Neither phenotype is shared by PCP genes, but both are highly reminiscent of what is observed following perturbation of centriolar or ciliary components (27,28). A closer examination of ciliary morphology in the chordotonal neurons responsible for mechanosensation in the animals' legs by DIC and immunofluorescence microscopy as well as ultrastructural analysis revealed only minor defects (fig.…”

“…4B,C). IFT and BBS proteins do not contribute to cilium biogenesis in Drosophila sperm (28,38) and were not detected.…”

In-cell structure and snapshots ofcopiaretrotransposons in intact tissue by cryo-electron tomography

“…The following fly lines were used throughout this study: w; tj-GAL4/CyO;; [64]; w* P{UAS-Sec31.RFP}13.5; snaSco/CyO, P{ftz-lacB}E3 (Bloomington #86533); emGFP::nup358 PBac{y[+mDint2] = vas-Cas9}VK00027 [40] ; w*; P{w+, sqhp>LifeAct::mNeonGreen}; MKRS/TM6. [65] (Gift from Dr. Cristina Pallares Cartes); w* P{UAS-Sec13::EosFP} / CyO; (this study); w; ; arrestin2/GD (VDRC# 20991); w;; vreteno/GD (VDRC# 34897); y[1] v[1]; P{y[+t7.7] v[+t1.8]=TRiP.HMS00004}attP2/TM3, Sb[1] (Bloomington #33613); P{w[+mC]=otu-GAL4::VP16.R}1,w[*];P{w[+mC]=GAL4-nanos.NGT}40; P{w[+mC]=GAL4::VP16-nanos.UTR} CG6325[MVD1]; (Bloomington #31777); w;; white sh; [48]; w; aub sh (w22, attP40, 2nd chrs); ago3 sh (v22, attP2 third chrs) [48]; w; NGT40 (2); bam-GAL4 (3) [31] and gift from Jeroen Dobbelaere [66].…”

“…The following fly lines were used throughout this study: w; tj-GAL4/CyO;; [64] (Bloomington #31777); w;; white sh; [48]; w; aub sh (w22, attP40, 2nd chrs); ago3 sh (v22, attP2 third chrs) [48]; w; NGT40 (2); bam-GAL4 (3) [31] and gift from Jeroen Dobbelaere [66].…”