Spatially controlled expression of the Drosophila pseudouridine synthase RluA-1

Wang, Chung-Chi; Lo, Jun-Chih; Chien, Cheng-Ting; Huang, Min-Lang

doi:10.1387/ijdb.103112cw

Cited by 7 publications

(8 citation statements)

References 20 publications

(19 reference statements)

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“…The Class IV samples showed high enrichment of ppk, Gr28b, and knot transcripts that are known to have enriched expression in these cells [1, 9, 10] (Figure 1A). As expected, the transcript for the pan-multidendritic neuron marker multiple [11] was seen in both Class IV and Class I samples. Remarkably, we found highly enriched expression in the Class IV neurons (relative to the Class I neurons) for a messenger RNA that encoded one of the ion channel subunits that was identified in our forward genetic screen ( CG8546 / ppk26 ) (Figure 1A).…”

Section: Resultssupporting

confidence: 73%

Balboa Binds to Pickpocket In Vivo and Is Required for Mechanical Nociception in Drosophila Larvae

et al. 2014

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Summary The Drosophila gene pickpocket (ppk) encodes an ion channel subunit of the Degenerin/Epithelial (DEG/ENaC) family [1]. PPK is specifically expressed in nociceptive, Class IV multidendritic (md) neurons and is functionally required for mechanical nociception responses [2, 3]. In this study, in a genome-wide genetic screen for other ion channel subunits required for mechanical nociception, we identify a gene that we name balboa (a.k.a. CG8546, ppk26) [4]. Interestingly, the balboa locus encodes a DEG/ENaC ion channel subunit highly similar in amino acid sequence to PPK [5]. Moreover, laser-capture isolation of RNA from larval neurons and microarray analyses reveal that balboa is also highly enriched in nociceptive neurons. The requirement for Balboa and PPK in mechanical nociception behaviors and their specific expression in larval nociceptors led us to hypothesize that these DEG/ENaC subunits form an ion channel complex in vivo. In nociceptive neurons Balboa∷GFP proteins distribute uniformly throughout dendrites but remarkably localize to discrete foci when ectopically expressed in other neuron subtypes (where PPK is not expressed). Indeed, ectopically co-expressing ppk transforms this punctate Balboa∷GFP expression pattern to the uniform distribution observed in its native cell-type. Furthermore, ppk-RNAi in Class IV neurons alters the broad Balboa∷GFP pattern to a punctate distribution. Interestingly, this interaction is mutually codependent as balboa-RNAi eliminates Venus∷PPK from the sensory dendrites of nociceptors. Finally, using a GFP-reconstitution approach in transgenic larvae, we directly detect in vivo physical interactions among PPK and Balboa subunits. Combined, our results indicate a critical mechanical nociception function for heteromeric PPK and Balboa channels in vivo.

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Section: Resultssupporting

confidence: 73%

Balboa Binds to Pickpocket In Vivo and Is Required for Mechanical Nociception in Drosophila Larvae

et al. 2014

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“…In Drosophila melanogaster , RluA enzymes modify uridines in rRNA and tRNA. In situ hybridization in embryos revealed a specific RluA-1 mRNA localization to dendrites of a subset of peripheral neurons, which also raises the question about the molecular function of RluA-1 and its target RNA in the peripheral nervous system during embryonic development (Wang et al, 2011 ).…”

Section: Pseudouridine (ψ)mentioning

confidence: 99%

The Emerging Field of Epitranscriptomics in Neurodevelopmental and Neuronal Disorders

Angelova

Dimitrova

Dinges

et al. 2018

Front. Bioeng. Biotechnol.

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Analogous to DNA methylation and histone modifications, RNA modifications represent a novel layer of regulation of gene expression. The dynamic nature and increasing number of RNA modifications offer new possibilities to rapidly alter gene expression upon specific environmental changes. Recent lines of evidence indicate that modified RNA molecules and associated complexes regulating and “reading” RNA modifications play key roles in the nervous system of several organisms, controlling both, its development and function. Mutations in several human genes that modify transfer RNA (tRNA) have been linked to neurological disorders, in particular to intellectual disability. Loss of RNA modifications alters the stability of tRNA, resulting in reduced translation efficiency and generation of tRNA fragments, which can interfere with neuronal functions. Modifications present on messenger RNAs (mRNAs) also play important roles during brain development. They contribute to neuronal growth and regeneration as well as to the local regulation of synaptic functions. Hence, potential combinatorial effects of RNA modifications on different classes of RNA may represent a novel code to dynamically fine tune gene expression during brain function. Here we discuss the recent findings demonstrating the impact of modified RNAs on neuronal processes and disorders.

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“…Previously described reporter genes for RluA-1 showed specific expression in larval multidendritic neurons (22). We replicated this finding by generating an RluA-1 GAL4 driver at the endogenous gene locus through recombination mediated cassette exchange (RMCE) of RluA-1 MI06897 (an intronic MiMIC) and a Trojan exon cassette (45).…”

Section: Resultsmentioning

confidence: 99%

“…RluA family members in bacteria are involved in ribosomal assembly and growth (43, 44) but their function in multicellular organisms has not been studied. Although pseudouridine synthases appear to function ubiquitously, as noted above, Drosophila RluA-1 , a member in RluA family, has been reported to be specifically expressed in md neurons (22). Thus, we have investigated the role for RluA-1 and its paralog RluA-2 , in nociception pathways that are known to depend on md neurons.…”

Section: Introductionmentioning

confidence: 96%

“…The historically first known genetic marker with specific expression pattern in the da neurons was the lacZ enhancer trap E7-2-36 (21). Later studies reported that this enhancer trap gene was inserted upstream of the RluA-1 gene and DNA sequences from upstream of RluA-1 caused expression of GAL4 in multidendritic neurons (22). The RluA1 gene encodes an enzyme that is predicted to have pseudouridine synthase activity, but how this RNA modifying protein is involved in the function of nociceptive multidendritic neurons remains unknown.…”

Section: Introductionmentioning

confidence: 99%

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Loss of pseudouridine synthases in the RluA family causes hypersensitive nociception inDrosophila

Song

Tracey

2019

Preprint

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Nociceptive neurons of Drosophila melanogaster larvae are characterized by highly branched dendritic processes whose proper morphogenesis relies on a large number of RNA-binding proteins. Post-transcriptional regulation of RNA in these dendrites has been found to play an important role in their function. Here, we investigate the neuronal functions of two putative RNA modification genes, RluA-1 and RluA-2, which are predicted to encode pseudouridine synthases. RluA-1 is specifically expressed in larval sensory neurons while RluA-2 expression is ubiquitous. Nociceptor-specific RNAi knockdown of RluA-1 caused hypersensitive nociception phenotypes, which were recapitulated with genetic null alleles. These were rescued with genomic duplication and nociceptor-specific expression of UAS-RluA-1-cDNA. As with RluA-1, RluA-2 loss of function mutants also displayed hyperalgesia. Interestingly, nociceptor neuron dendrites showed a hyperbranched morphology in the RluA-1 mutants. The latter may be a cause or a consequence of heightened sensitivity in mutant nociception behaviors.Author SummaryPseudouridine (Psi) is a C5-glycoside isomer of uridine and it is the most common posttranscriptional modification of RNAs, including noncoding tRNAs, rRNAs, snRNAs as well as mRNAs. Although first discovered in the 1950s, the biological functions of Psi in multicellular organisms are not well understood. Interestingly, a marker for sensory neurons in Drosophila encodes for a putative pseudouridine synthase called RluA-1. Here, we report our characterization of nociception phenotypes for larvae with RluA-1 loss of function along with that of a related gene RluA-2. Disrupting either or both RluA-1 and RluA-2 resulted in hypersensitive nociception. In addition, RluA-1 mutants have more highly branched nociceptor neurites that innervate the epidermis. Our studies suggest an important role for the RluA family in nociception. This may be through its action on RNAs that regulate neuronal excitability and/or dendrite morphogenesis.

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Spatially controlled expression of the Drosophila pseudouridine synthase RluA-1

Cited by 7 publications

References 20 publications

Balboa Binds to Pickpocket In Vivo and Is Required for Mechanical Nociception in Drosophila Larvae

Balboa Binds to Pickpocket In Vivo and Is Required for Mechanical Nociception in Drosophila Larvae

The Emerging Field of Epitranscriptomics in Neurodevelopmental and Neuronal Disorders

Loss of pseudouridine synthases in the RluA family causes hypersensitive nociception inDrosophila

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