Spatial and temporal expression of dADAR mRNA and protein isoforms during embryogenesis in Drosophila melanogaster

Chen, Jing; Lakshmi, G. Girija; Hays, Danielle L.; McDowell, Katherine M.; Ma, Enbo; Vaughn, Jack C.

doi:10.1016/j.diff.2009.08.003

Cited by 8 publications

(41 citation statements)

References 27 publications

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“…The function of Hsc70Cb, into which KP70C is inserted, is speculated to assist protein folding, and its expression is positive in embryonic and adult stages (Flybase 2004). The lack of detection of RNA editing in the adult stage in this study is coincident with observations that expression of dADAR is limited in the central nervous system after the early embryonic stage (Palladino et al 2000a, b;Ma et al 2001;Chen et al 2009). …”

Section: Discussionsupporting

confidence: 90%

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RNA editing in P transposable element read-through transcripts in Drosophila melanogaster

Fukui

Itoh

2010

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RNA editing is proposed as a modulator of transcriptomes, but its biological impact has not been fully elucidated. In particular, its importance for transposable elements is controversial. We found RNA editing on antisense read-through transcripts of KP elements, one of the deletion derivatives of P transposable elements in Drosophila melanogaster. Three kinds of RNA editing were detected at 20 sites around the terminal inverted repeats (TIR); 15 A-to-G, four U-to-C, and one C-to-U conversions. A-to-G conversions are suggested to be attributed to A-to-I RNA editing on KP element RNAs, because inosine (I) in RNA is recognized as G by reverse transcriptase. TIRs were deduced to form dsRNAs as a putative target of ADAR. This is the first report of RNA editing on mobile elements of Drosophila.

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Section: Discussionsupporting

confidence: 90%

“…Recent observation of a ubiquitous distribution of dADAR mRNA in embryos (Chen et al 2009) supports this assumption. It is noteworthy that KP22B and KP70C are the only two KP elements oppositely inserted into introns among the 16 P elements examined.…”

Section: Discussionmentioning

confidence: 86%

RNA editing in P transposable element read-through transcripts in Drosophila melanogaster

Fukui

Itoh

2010

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“…2). Preliminary studies have shown that the abundance of the longer rnp-4f mRNA isoform in dADAR null mutant is about 30% less than that seen in wild-type during embryogenesis (Chen et al, 2009), suggesting a role for dADAR protein in the intron 0 alternative splicing decision. Taken together, these observations led us to hypothesize that dADAR protein may target rnp-4f transcripts within the 5′-UTR and regulate alternative splicing.…”

Section: Introductionmentioning

confidence: 99%

“…Insofar as the major difference between the two mRNA isoforms is found in their differing 5′-UTRs, our attention in this report is focused on this region to understand the regulation of the splicing event. The longer isoform contains a highly conserved secondary structure in which intron 0 pairs with part of adjacent exon 2 (Chen et al, 2009), itself highly conserved, to form a long stable stem-loop (Fig. 1).…”

Section: Introductionmentioning

confidence: 99%

“…dADAR encodes a protein which utilizes double-stranded RNAs for substrate in the A-to-I editing of some pre-mRNAs (reviewed in Bass, 2001; Gott, 2007). The single-copy dADAR gene encodes two different major mRNA isoforms, designated full-length (Palladino et al, 2000) and truncated (Ma et al, 2002; Chen et al, 2009), of which both contain a double-stranded RNA binding motif but only the longer variant contains a catalytic domain (Fig. 2).…”

Section: Introductionmentioning

confidence: 99%

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An RNA electrophoretic mobility shift and mutational analysis of rnp-4f 5′-UTR intron splicing regulatory proteins in Drosophila reveals a novel new role for a dADAR protein isoform

Lakshmi

Ghosh

Jones

et al. 2012

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Alternative splicing greatly enhances the diversity of proteins encoded by eukaryotic genomes, and is also important in gene expression control. In contrast to the great depth of knowledge as to molecular mechanisms in the splicing pathway itself, relatively little is known about the regulatory events behind this process. The 5′-UTR and 3′-UTR in pre-mRNAs play a variety of roles in controlling eukaryotic gene expression, including translational modulation, and nearly 4,000 of the roughly 14,000 protein coding genes in Drosophila contain introns of unknown functional significance in their 5′-UTR. Here we report the results of an RNA electrophoretic mobility shift analysis of Drosophila rnp-4f 5′-UTR intron 0 splicing regulatory proteins. The pre-mRNA potential regulatory element consists of an evolutionarily-conserved 177-nt stem-loop arising from pairing of intron 0 with part of adjacent exon 2. Incubation of in vitro transcribed probe with embryo protein extract is shown to result in two shifted RNA-protein bands, and protein extract from a dADAR null mutant fly line results in only one shifted band. A mutated stem-loop in which the conserved exon 2 primary sequence is changed but secondary structure maintained by introducing compensatory base changes results in diminished band shifts. To test the hypothesis that dADAR plays a role in intron splicing regulation in vivo, levels of unspliced rnp-4f mRNA in dADAR mutant were compared to wild-type via real-time qRT-PCR. The results show that during embryogenesis unspliced rnp-4f mRNA levels fall by up to 85% in the mutant, in support of the hypothesis. Taken together, these results demonstrate a novel role for dADAR protein in rnp-4f 5′-UTR alternative intron splicing regulation which is consistent with a previously proposed model.

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5′-UTR mediated translational control of splicing assembly factor RNP-4F expression during development of the Drosophila central nervous system

Chen

Yang

Doctor

et al. 2013

Gene

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Spatial and temporal expression of dADAR mRNA and protein isoforms during embryogenesis in Drosophila melanogaster

Cited by 8 publications

References 27 publications

RNA editing in P transposable element read-through transcripts in Drosophila melanogaster

RNA editing in P transposable element read-through transcripts in Drosophila melanogaster

An RNA electrophoretic mobility shift and mutational analysis of rnp-4f 5′-UTR intron splicing regulatory proteins in Drosophila reveals a novel new role for a dADAR protein isoform

5′-UTR mediated translational control of splicing assembly factor RNP-4F expression during development of the Drosophila central nervous system

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