Ciliary and non-ciliary expression and function of PACRGduring vertebrate development

Thumberger, Thomas; Hagenlocher, Cathrin; Tisler, Matthias; Beyer, Tina; Tietze, Nina; Schweickert, Axel; Feistel, Kerstin; Blum, Martin

doi:10.1186/2046-2530-1-13

Cited by 16 publications

(17 citation statements)

References 40 publications

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“…Due to its axonemal functions, knockdown of PACRG genes in Trypanosoma brucei and Xenopus , lead to defects in the doublet structure and, therefore, impaired flagellar motility. In vertebrates, defects in left-right body symmetry, neural tube closure were observed from knockdowns of PACRG (19). In mice, PACRG knockout results in male sterility (20) and hydrocephalus (21).…”

Section: Introductionmentioning

confidence: 99%

The inner junction complex of the cilia is an interaction hub that involves tubulin post-translational modifications

Khalifa

Ichikawa

Dai

et al. 2019

Preprint

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Microtubules are cytoskeletal structures involved in structural support, microtubule-based transport and the organization of organelles in the cells. The building blocks of the microtubule, the α- and β-tubulin heterodimers, polymerize into protofilaments, that associate laterally to form the hollow microtubule. There exists a specific type of microtubule structures in the cilia, termed doublet microtubules, where high stability is required for ciliary beating and function. The doublet microtubule, consisting of a complete A-tubule and a partial B-tubule maintains its stability through unique interactions at its outer and inner junctions, where the A- and B-tubules meet.Using cryo-electron microscopy, we present the answer to the long-standing question regarding the identities, localizations and structures of the Chlamydomonas doublet microtubule inner junction proteins. Using a combination of sequence bioinformatics and mass spectrometry, we identified two new inner junction proteins, FAP276 and FAP106, and an inner junction associated protein FAP126. We show that inner junction proteins PACRG and FAP20, together with FAP52, previously unidentified FAP276, FAP106 and FAP126, form an interaction hub at the inner junction, which involves tubulin sites for post-translational modifications. We further compare the Chlamydomonas and Tetrahymena doublet microtubule structures to understand the common and species-specific features of the inner junction.

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Section: Introductionmentioning

confidence: 99%

The inner junction complex of the cilia is an interaction hub that involves tubulin post-translational modifications

Khalifa

Ichikawa

Dai

et al. 2019

Preprint

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“…PACRG has been localized to the axoneme (Dawe et al ., 2005; Lechtreck et al ., 2009; Thumberger et al ., 2012). Loss of PACRG results in male sterility in mice (Lorenzetti et al ., 2004) as well as defects in ependymal ciliary motility and hydrocephalus (Wilson et al ., 2009; Wilson et al ., 2010).…”

Section: Introductionmentioning

confidence: 99%

“…PACRG was originally identified in the human genome as a gene, whose transcription is coregulated with a Parkinson's disease-related gene Parkin by a bi-directional promoter (Kitada et al, 1998;West et al, 2003). PACRG has been localized to the axoneme (Dawe et al, 2005;Lechtreck et al, 2009;Thumberger et al, 2012). Loss of PACRG results in male sterility in mice (Lorenzetti et al, 2004) as well as defects in ependymal ciliary motility and hydrocephalus (Wilson et al, 2009;Wilson et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

Microtubule binding protein PACRG plays a role in regulating specific ciliary dyneins during microtubule sliding

Mizuno¹,

Dymek

Smith

2016

Cytoskeleton

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The complex waveforms characteristic of motile eukaryotic cilia and flagella are produced by the temporally and spatially regulated action of multiple dynein subforms generating sliding between subsets of axonemal microtubules. Multiple protein complexes have been identified that are associated with the doublet microtubules and that mediate regulatory signals between key axonemal structures, such as the radial spokes and central apparatus, and the dynein arm motors; these complexes include the N-DRC, MIA, and CSC complexes. Previous studies have shown that PACRG (parkin co-regulated gene) forms a complex that is anchored to the axonemal doublet microtubules. Loss of PACRG causes defects in ciliary motility and cilia related diseases. Here, we use an in vitro microtubule sliding assay to demonstrate that PACRG and its interactors are part of a signaling pathway that includes the central apparatus, radial spokes and specific inner dynein arm subforms to control dynein-driven microtubule sliding. Using a biochemical approach, our studies also indicate that PACRG interacts with the radial spokes.

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“…SNAP2, on the other hand, uses evolutionary, structural, solvent-PARK2)[63]. PACRG is affiliated with axonemal doublet microtubules, and contributes to the signaling pathway that controls dynein-driven microtubule sliding[62,[64][65][66]. A single nucleotide variant (SNV) in PACRG was strongly associated with an increased risk of developing childhood asthma following early-life tobacco smoke exposure[67].For the RTTN substitution variant, tryptophan is an aromatic, non-polar and hydrophobic amino acid often buried in hydrophobic cores, while arginine is a polar and positively charged amino acid often found on outside chains[68].…”

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confidence: 99%

Peer Review #1 of "Sequence variant analysis of RNA sequences in severe equine asthma (v0.2)"

2018

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Background. Severe equine asthma is a chronic inflammatory disease of the lung in horses similar to low-Th2 late-onset asthma in humans. This study aimed to determine the utility of RNA-Seq to call gene sequence variants, and to identify sequence variants or potential relevance to the pathogenesis of asthma. Methods. RNA-Seq data were generated from endobronchial biopsies collected from 6 asthmatic and 7 non-asthmatic horses before and after challenge (26 samples total). Sequences were aligned to the equine genome with Spliced Transcripts Alignment to Reference software. Read preparation for sequence variant calling was performed with Picard tools and Genome Analysis Toolkit (GATK). Sequence variants were called and filtered using GATK and Ensembl Variant Effect Predictor (VEP) tools, and two RNA-Seq predicted sequence variants were investigated with both PCR and Sanger sequencing. Supplementary analysis of novel sequence variant selection with VEP was based on a score of <0.01 predicted with Sorting Intolerant From Tolerant (SIFT) software, missense nature, location within the protein coding sequence and presence in all asthmatic individuals. For select variants, effect on protein function was assessed with Polymorphism Phenotyping (PolyPhen) 2 and Screening for Non-Acceptable Polymorphism (SNAP) 2 software. Sequences were aligned and 3D protein structures predicted with Geneious software. Difference in allele frequency between the groups was assessed using a Pearson's Chi-squared test with Yates' continuity correction, and difference in genotype frequency was calculated using the Fisher's exact test for count data. Results. RNA-Seq variant calling and filtering correctly identified substitution variants in PACRG and RTTN. Sanger sequencing confirmed that the PACRG substitution was appropriately identified in all 26 samples while the RTTN substitution was identified correctly in 24 of 26 samples. These variants of uncertain significance had substitutions that were predicted to result in loss of function and to be non-neutral. Amino acid substitutions projected no change of hydrophobicity and

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Ciliary and non-ciliary expression and function of PACRGduring vertebrate development

Cited by 16 publications

References 40 publications

The inner junction complex of the cilia is an interaction hub that involves tubulin post-translational modifications

The inner junction complex of the cilia is an interaction hub that involves tubulin post-translational modifications

Microtubule binding protein PACRG plays a role in regulating specific ciliary dyneins during microtubule sliding

Peer Review #1 of "Sequence variant analysis of RNA sequences in severe equine asthma (v0.2)"

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