Translational Control by the DEAD Box RNA Helicase belle Regulates Ecdysone-Triggered Transcriptional Cascades

Ihry, Robert J.; Sapiro, Anne L.; Bashirullah, Arash

doi:10.1371/journal.pgen.1003085

Cited by 43 publications

(68 citation statements)

References 43 publications

(62 reference statements)

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“…mRNA was extracted from 1 to 3 days old flies using RNAeasy (Qiagen, Hilden, Germany) and quantitative polymerase chain reaction (qPCR) was performed with the following dronc-specific primers: Nedd2-like caspase forward primer (NcF) 5'-CTCGCTAAACGAACGGAGAAC-3' and Nedd2-like caspase reverse primer (NcR) 5'-CAACGACACCCACATAAGGG-3', as described. 79 Tubulin was used for normalization.…”

Section: Methodsmentioning

confidence: 99%

The initiator caspase Dronc is subject of enhanced autophagy upon proteasome impairment in Drosophila

et al. 2016

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A major function of ubiquitylation is to deliver target proteins to the proteasome for degradation. In the apoptotic pathway in Drosophila, the inhibitor of apoptosis protein 1 (Diap1) regulates the activity of the initiator caspase Dronc (death regulator Nedd2-like caspase; caspase-9 ortholog) by ubiquitylation, supposedly targeting Dronc for degradation by the proteasome. Using a genetic approach, we show that Dronc protein fails to accumulate in epithelial cells with impaired proteasome function suggesting that it is not degraded by the proteasome, contrary to the expectation. Similarly, decreased autophagy, an alternative catabolic pathway, does not result in increased Dronc protein levels. However, combined impairment of the proteasome and autophagy triggers accumulation of Dronc protein levels suggesting that autophagy compensates for the loss of the proteasome with respect to Dronc turnover. Consistently, we show that loss of the proteasome enhances endogenous autophagy in epithelial cells. We propose that enhanced autophagy degrades Dronc if proteasome function is impaired.

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Section: Methodsmentioning

confidence: 99%

The initiator caspase Dronc is subject of enhanced autophagy upon proteasome impairment in Drosophila

et al. 2016

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“…Images of dissected pupae and of persistent salivary glands were captured using an Olympus SZX16 stereomicroscope coupled to a digital camera. Staging and heat-shock treatments for ectopic expression of E74A were performed with control and mutant animals carrying one copy of the hs-E74A transgene as previously described (Ihry et al 2012).…”

Section: Developmental Staging and Psg Phenotypesmentioning

confidence: 99%

“…To quantify messenger RNA (mRNA) of target genes in whole animals and dissected salivary glands, quantitative real-time reverse transcriptase PCR (qPCR) was performed as previously described (Ihry et al 2012). RNA was isolated using the RNeasy Plus Mini kit (QIAGEN, Valencia, CA).…”

Section: Quantitative Rt-pcrmentioning

confidence: 99%

“…In all cases, experimental samples and matched controls were analyzed simultaneously for expression of target genes and the reference gene rp49. Primer sequences for reaper, hid, Nc, Ark, E74A, E75A, E93, and rp49 were previously described (Ihry et al 2012); the remaining primers used in this study are listed in Supporting Information, Table S1. Relative expression was calculated using the Relative Expression Software Tool (REST) (Pfaffl et al 2002) as previously described (Ihry et al 2012).…”

Section: Quantitative Rt-pcrmentioning

confidence: 99%

“…Primer sequences for reaper, hid, Nc, Ark, E74A, E75A, E93, and rp49 were previously described (Ihry et al 2012); the remaining primers used in this study are listed in Supporting Information, Table S1. Relative expression was calculated using the Relative Expression Software Tool (REST) (Pfaffl et al 2002) as previously described (Ihry et al 2012). Each data point used represents the analysis of three independent biological samples (except as indicated in Figure 1B).…”

Section: Quantitative Rt-pcrmentioning

confidence: 99%

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Genetic Control of Specificity to Steroid-Triggered Responses in Drosophila

Ihry

Bashirullah

2014

Genetics

Self Cite

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Steroid hormones trigger a wide variety of biological responses through stage-and tissue-specific activation of target gene expression. The mechanisms that provide specificity to systemically released pulses of steroids, however, remain poorly understood. We previously completed a forward genetic screen for mutations that disrupt the destruction of larval salivary glands during metamorphosis in Drosophila melanogaster, a process triggered by the steroid hormone 20-hydroxyecdysone (ecdysone). Here, we characterize 10 complementation groups mapped to genes from this screen. Most of these mutations disrupt the ecdysone-induced expression of death activators, thereby failing to initiate tissue destruction. However, other responses to ecdysone, even within salivary glands, occur normally in mutant animals. Many of these newly identified regulators of ecdysone signaling, including brwd3, med12, med24, pak, and psg2, represent novel components of the ecdysone-triggered transcriptional hierarchy. These genes function combinatorially to provide specificity to ecdysone pulses, amplifying the hormonal cue in a stage-, tissue-, and target gene-specific manner. Most of the ecdysone response genes identified in this screen encode homologs of mammalian nuclear receptor coregulators, demonstrating an unexpected degree of functional conservation in the mechanisms that regulate steroid signaling between insects and mammals.

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The RNA helicase DDX3 is required for ovarian development and oocyte maturation in Locusta migratoria

Wang

Deng

et al. 2021

Arch Insect Biochem Physiol

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DDX3 represents a well‐defined subfamily of DEAD‐box RNA helicase and exerts multiple functions in RNA metabolism, cell cycle, tumorigenesis, signal pathway, and fertility. Our previous study has shown that LmDDX3, the ortholog of DDX3 in Locusta migratoria, is ubiquitously expressed, and with a high abundance in testis and ovary. Knockdown of LmDDX3 results in a lethal phenotype in nymph, but it still remains unclear for its role in reproductive process. In this study, we therefore characterized LmDDX3 expression in female adult locust and analyzed its function in oocyte development. LmDDX3 was expressed in all tissues examined with significant more transcripts in ovary and hindgut. In ovary, a strong expression level was detected at the day just after adult eclosion, and a dramatic reduction then occurred during the oocyte development. LmDDX3 RNAi led to a reduced vitellogenin (Vg) expression in fat body via partially at least, the JH signaling pathway, and caused an upregulation of vitellogenin receptor (VgR) in ovary, and thus blocked the ovarian development and oocyte maturation. Sequence and phylogenetic analysis indicated that LmDDX3 was closely related to termite DDX3. Taken together, these data reveal a critical role for LmDDX3 in regulating the transcription of Vg and VgR, two major factors in vitellogenesis that is a key process required for ovary development and oocyte maturation in locust, and contribute thereof a new putative target for locust biological control.

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Translational Control by the DEAD Box RNA Helicase belle Regulates Ecdysone-Triggered Transcriptional Cascades

Cited by 43 publications

References 43 publications

The initiator caspase Dronc is subject of enhanced autophagy upon proteasome impairment in Drosophila

The initiator caspase Dronc is subject of enhanced autophagy upon proteasome impairment in Drosophila

Genetic Control of Specificity to Steroid-Triggered Responses in Drosophila

The RNA helicase DDX3 is required for ovarian development and oocyte maturation in Locusta migratoria

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