Joint action of two RNA degradation pathways controls the timing of maternal transcript elimination at the midblastula transition in Drosophila melanogaster

Bashirullah, Arash; Halsell, Susan R.; Cooperstock, Ramona L.; Kloc, Małgorzata; Karaiskakis, Angelo; Fisher, William W.; Fu, Weili; Hamilton, Jill; Etkin, Laurence D.; Lipshitz, Howard D.

doi:10.1093/emboj/18.9.2610

Cited by 201 publications

(269 citation statements)

References 42 publications

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“…2a). Regulatory elements of these genes are known to cause germ cell-specific expression, either by specifically directing zygotic transcription in these cells (vasa) (16) or by spatially regulating translation and stability of maternal mRNA, which leads to expression at the posterior of the embryo where germ cells form (nanos) (17)(18)(19)(20)(21)(22)(23).…”

Section: Resultsmentioning

confidence: 99%

An optimized transgenesis system for Drosophila using germ-line-specific φC31 integrases

Bischof

Maeda²,

Hediger³

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

1,826

2,089

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Germ-line transformation via transposable elements is a powerful tool to study gene function in Drosophila melanogaster. However, some inherent characteristics of transposon-mediated transgenesis limit its use for transgene analysis. Here, we circumvent these limitations by optimizing a C31-based integration system. We generated a collection of lines with precisely mapped attP sites that allow the insertion of transgenes into many different predetermined intergenic locations throughout the fly genome. By using regulatory elements of the nanos and vasa genes, we established endogenous sources of the C31 integrase, eliminating the difficulties of coinjecting integrase mRNA and raising the transformation efficiency. Moreover, to discriminate between specific and rare nonspecific integration events, a white gene-based reconstitution system was generated that enables visual selection for precise attP targeting. Finally, we demonstrate that our chromosomal attP sites can be modified in situ, extending their scope while retaining their properties as landing sites. The efficiency, ease-of-use, and versatility obtained here with the C31-based integration system represents an important advance in transgenesis and opens up the possibility of systematic, highthroughput screening of large cDNA sets and regulatory elements.attP landing sites ͉ germ-line transformation ͉ site-specific integration A major goal in the present era of genomics is to identify and functionally characterize all genes relevant to a specific pathway or biological process. With its powerful repertoire of genetic tools, the multicellular model organism Drosophila melanogaster has played an eminent role in this endeavor (1). One method to identify relevant genes is to perform chemical mutagenesis screens of various kinds. Another very fruitful approach in Drosophila has been the use of P-element-mediated germ-line transformation (2, 3), especially when combined with tools such as the Gal4/UAS expression system (4) or when it is used for insertional mutagenesis (5, 6). One characteristic of P-elements is their random integration behavior. Although this ''randomness'' is advantageous for generating mutations and deletions, it is generally not ideal for transgene analysis. The random integration of P-elements necessitates considerable effort to map insertions. Genomic position effects complicate the analysis of transgenes and render precise structure/ function analyses nearly impossible. A further shortcoming of the P-element system is its relatively moderate transformation efficiency, a significant hurdle to any large-scale transgenesis effort.Strategies have been developed to circumvent the problem of randomness by targeted integration systems in Drosophila, which are generally based on the FLP and Cre recombinases (7-9, 32). Such techniques permit precise targeting to genomic landing sites but are still handicapped by transformation rates that are, at best, moderately higher than those achieved with the P-element system (8). Furthermore, especially for FL...

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Section: Resultsmentioning

confidence: 99%

An optimized transgenesis system for Drosophila using germ-line-specific φC31 integrases

Bischof

Maeda²,

Hediger³

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

1,826

2,089

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“…For instance, in Drosophila, the maternal nanos message is distributed uniformly throughout the cytoplasm of the mature oocyte. Upon activation nanos mRNA is degraded everywhere except the posterior pole of the oocyte, where it is translated and functions in posterior patterning (Bashirullah et al, 1999;Bergsten and Gavis, 1999). Degradation can also remove mRNAs whose products are no longer needed or that interfere with later development, such as during the maternal to zygotic transition (MZT; mammals, zebrafish) or mid-blastula transition (MBT; Xenopus, Drosophila) (Bashirullah et al, 1999;Hamatani et al, 2004;Giraldez et al, 2006;Tadros et al, 2007).…”

Section: Maternal Transcript Degradationmentioning

confidence: 99%

Transitioning from egg to embryo: Triggers and mechanisms of egg activation

Horner

Wolfner

2008

Developmental Dynamics

183

177

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The transition from mature oocyte to developing embryo requires a coordinated series of events, collectively known as egg activation. Egg activation includes changes to egg coverings to prevent polyspermy, release of oocyte meiotic arrest, generation of haploid female and male pronuclei, changes in maternal mRNAs and protein populations, and cytoskeletal rearrangements. In many animals, egg activation is triggered by fertilization, which increases intracellular calcium within the oocyte and thereby regulates molecular events of egg activation. In other animals, fertilization-independent external signals, including mechanical stimulation of eggs and/or changes in ionic milieu, trigger activation. Recent studies have clarified the upstream portion of pathways leading to eggshell changes and cell cycle resumption and have identified activation-induced changes in maternal mRNA and protein profiles that can identify molecular players in the downstream events of egg activation. We review signals that trigger activation and how they link to subsequent molecular events of egg activation. Developmental Dynamics 237:527-544, 2008.

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“…In the Drosophila embryo, hsp83 and nanos mRNA are degraded throughout the embryo except for the posterior pole where they are protected from degradation by elements in their 3 0 UTR. 63,64 Second, mRNAs passively diffusing through the cytoplasm can be bound and retained by anchoring proteins that are themselves localized to specific cellular sites. This system has been shown to function in at least two different systems: Xenopus oocytes 65 with the RNAs Xcat2 and Xdazl and Drosophila embryos with nanos mRNA.…”

Section: Protein Factors Involved In Rna Localizationmentioning

confidence: 99%

Genomic analysis of RNA localization

2014

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The localization of mRNAs to specific subcellular sites is widespread, allowing cells to spatially restrict and regulate protein production, and playing important roles in development and cellular physiology. This process has been studied in mechanistic detail for several RNAs. However, the generality or specificity of RNA localization systems and mechanisms that impact the many thousands of localized mRNAs has been difficult to assess. In this review, we discuss the current state of the field in determining which RNAs localize, which RNA sequences mediate localization, the protein factors involved, and the biological implications of localization. For each question, we examine prominent systems and techniques that are used to study individual messages, highlight recent genome-wide studies of RNA localization, and discuss the potential for adapting other high-throughput approaches to the study of localization.

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Joint action of two RNA degradation pathways controls the timing of maternal transcript elimination at the midblastula transition in Drosophila melanogaster

Cited by 201 publications

References 42 publications

An optimized transgenesis system for Drosophila using germ-line-specific φC31 integrases

An optimized transgenesis system for Drosophila using germ-line-specific φC31 integrases

Transitioning from egg to embryo: Triggers and mechanisms of egg activation

Genomic analysis of RNA localization

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