Novel cis-regulatory regions in ecdysone responsive genes are sufficient to promote gene expression in Drosophila ovarian cells

McDonald, Samantha I; Beachum, Allison N.; Hinnant, Taylor D.; Blake, Amelia J.; Bynum, Tierra; Hickman, E. Parris; Barnes, Joseph; Churchill, Kaely L.; Roberts, Tamesia S.; Zangwill, Denise E.; Ables, Elizabeth T.

doi:10.1016/j.gep.2019.119074

Cited by 5 publications

(5 citation statements)

References 63 publications

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“…For assessment of Ftz-f1 expression in the ovary, we used transgenic line ftz-f1 sfGFP, Tag: FLAG ( ftz-f1::GFP ), which carries a bacterial artificial chromosome containing the entire ftz-f1 gene locus and surrounding regulatory DNA, including an sfGFP-Tag: FLAG cassette introduced at the C-terminal end of the ftz-f1 coding region (Bloomington stock #38645; R. Spokony). Expression of ftz-f1 in escort cells and follicle cells was further confirmed using P{VT032964-GAL4}attP2 ( VT032964-Gal4 ) and P{VT032969-GAL4}attP2 ( VT032969-Gal4 ), in which ∼2 kB of intronic sequence from ftz-f1-RB is fused upstream of a Drosophila synthetic core promoter and GAL4 ( Kvon et al 2014 ; McDonald et al 2019 ). VT032964-Gal4 and VT032969-Gal4 were crossed with P{w+mC=UAS-lacZ.NZ}J312 ( UASt-lacZ ; Bloomington stock #3956) to confirm driver expression.…”

Section: Methodsmentioning

confidence: 87%

“…This pattern is consistent with a recent study in which high levels of Ftz-f1 protein were detected transiently in follicle cells in stages 10–12 ( Knapp et al 2020 ). To further validate the ftz-f1 expression in early follicle cells, we used two ftz-f1 enhancer trap lines ( ftz-f1 VT032964 and ftz-f1 VT032969 ) that are sufficient to drive reporter gene expression in the ovary ( McDonald et al 2019 ). Both reporters correspond to α- ftz-f1 intronic DNA, and ftz-f1 VT032964 overlaps a previously characterized β- ftz-f1 enhancer element ( Kageyama et al 2003 ; McDonald et al 2019 ).…”

Section: Resultsmentioning

confidence: 99%

“…To further validate the ftz-f1 expression in early follicle cells, we used two ftz-f1 enhancer trap lines ( ftz-f1 VT032964 and ftz-f1 VT032969 ) that are sufficient to drive reporter gene expression in the ovary ( McDonald et al 2019 ). Both reporters correspond to α- ftz-f1 intronic DNA, and ftz-f1 VT032964 overlaps a previously characterized β- ftz-f1 enhancer element ( Kageyama et al 2003 ; McDonald et al 2019 ). Similar to Ftz-f1::GFP expression, ftz-f1 enhancer trap lines were able to drive expression of a lacZ reporter in cap cells and escort cells in the germarium ( ftz-f1 VT032964 ; Figure 1E ) and some main body follicle cells in stages 5–10B (both reporters; Figure 1, F and G ).…”

Section: Resultsmentioning

confidence: 99%

“…Unfortunately, it has been difficult to tease apart the interactions between these factors, given the high degree of egg chamber death in loss-of-function mutants. Our recent identification of cis-regulatory enhancers located in ecdysone response genes ( McDonald et al 2019 ) may prove to be useful reagents to analyze these complex genetic interactions.…”

Section: Discussionmentioning

confidence: 99%

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Orphan nuclear receptorftz-f1 (NR5A3)promotes egg chamber survival in theDrosophilaovary

Beachum

Whitehead

McDonald

et al. 2021

G3 Genes|Genomes|Genetics

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Gamete production in mammals and insects is controlled by cell signaling pathways that facilitate communication between germ cells and somatic cells. Nuclear receptor signaling is a key mediator of many aspects of reproduction, including gametogenesis. For example, the NR5A sub-family of nuclear receptors are essential for gonad development and sex steroid production in mammals. Despite the original identification of the NR5A sub-family in the model insect Drosophila melanogaster, it has been unclear whether Drosophila NR5A receptors directly control oocyte production. Ftz-f1 is expressed throughout the ovary, including in germline stem cells, germline cysts, and several populations of somatic cells. We demonstrate that ftz-f1 is required in follicle cells prior to stage 10 to promote egg chamber survival at the mid-oogenesis checkpoint. Our data suggest that egg chamber death in the absence of ftz-f1 is due, at least in part, to failure of follicle cells to exit the mitotic cell cycle or failure to accumulate oocyte-specific factors in the germline. Taken together, these results demonstrate that, as in mammals, the NR5A sub-family promotes maximal reproductive output in Drosophila. Our data underscore the importance of nuclear receptors in the control of reproduction and highlight the utility of Drosophila oogenesis as a key model for unraveling the complexity of nuclear receptor signaling in gametogenesis.

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Section: Methodsmentioning

confidence: 87%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

See 2 more Smart Citations

Orphan nuclear receptorftz-f1 (NR5A3)promotes egg chamber survival in theDrosophilaovary

Beachum

Whitehead

McDonald

et al. 2021

G3 Genes|Genomes|Genetics

View full text Add to dashboard Cite

show abstract

“…PriA represents a striking case of a pleiotropic enhancer that is expressed in both the embryonic epidermis and the leg primordium, as well as in renal (Bohère et al, 2018) and intestinal (Al Hayek et al, 2021) adult stem cells. Of note, other genes of the ecdysone cascade possess pleiotropic enhancers, as seen with EcR, Eip75B, and ftz-f1 that are driven by enhancers active in both somatic and germline tissues during oogenesis (McDonald et al, 2019). Pleiotropic enhancers may serve as a platform to drive gene expression in several spatiotemporal contexts (Sabaris et al, 2019), and future work will be required to better understand their behavior and functions during development (Preger-Ben Noon et al, 2018).…”

Section: The Pri Gene Displays Hallmarks Of a Master Developmental Genementioning

confidence: 99%

Pri smORF Peptides Are Wide Mediators of Ecdysone Signaling, Contributing to Shape Spatiotemporal Responses

et al. 2021

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There is growing evidence that peptides encoded by small open-reading frames (sORF or smORF) can fulfill various cellular functions and define a novel class regulatory molecules. To which extend transcripts encoding only smORF peptides compare with canonical protein-coding genes, yet remain poorly understood. In particular, little is known on whether and how smORF-encoding RNAs might need tightly regulated expression within a given tissue, at a given time during development. We addressed these questions through the analysis of Drosophila polished rice (pri, a.k.a. tarsal less or mille pattes), which encodes four smORF peptides (11–32 amino acids in length) required at several stages of development. Previous work has shown that the expression of pri during epidermal development is regulated in the response to ecdysone, the major steroid hormone in insects. Here, we show that pri transcription is strongly upregulated by ecdysone across a large panel of cell types, suggesting that pri is a core component of ecdysone response. Although pri is produced as an intron-less short transcript (1.5 kb), genetic assays reveal that the developmental functions of pri require an unexpectedly large array of enhancers (spanning over 50 kb), driving a variety of spatiotemporal patterns of pri expression across developing tissues. Furthermore, we found that separate pri enhancers are directly activated by the ecdysone nuclear receptor (EcR) and display distinct regulatory modes between developmental tissues and/or stages. Alike major developmental genes, the expression of pri in a given tissue often involves several enhancers driving apparently redundant (or shadow) expression, while individual pri enhancers can harbor pleiotropic functions across tissues. Taken together, these data reveal the broad role of Pri smORF peptides in ecdysone signaling and show that the cis-regulatory architecture of the pri gene contributes to shape distinct spatial and temporal patterns of ecdysone response throughout development.

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Anatomy and evolution of a DNA replication origin

et al. 2021

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Novel cis-regulatory regions in ecdysone responsive genes are sufficient to promote gene expression in Drosophila ovarian cells

Cited by 5 publications

References 63 publications

Orphan nuclear receptorftz-f1 (NR5A3)promotes egg chamber survival in theDrosophilaovary

Orphan nuclear receptorftz-f1 (NR5A3)promotes egg chamber survival in theDrosophilaovary

Pri smORF Peptides Are Wide Mediators of Ecdysone Signaling, Contributing to Shape Spatiotemporal Responses

Anatomy and evolution of a DNA replication origin

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