Dynein Associates with oskar mRNPs and Is Required For Their Efficient Net Plus-End Localization in Drosophila Oocytes

Sanghavi, Paulomi; Laxani, Shobha; Li, Xuan; Bullock, Simon L.; Gonsalvez, Graydon B.

doi:10.1371/journal.pone.0080605

Cited by 26 publications

(40 citation statements)

References 56 publications

(88 reference statements)

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“…In situ hybridization osk, bcd, and grk mRNAs were visualized as previously described (Sanghavi et al 2013).…”

Section: Immunofluorescencementioning

confidence: 99%

“…A total of 600 mg of each lysate was then incubated with GFPTrap beads (Chromotek) to purify GFP and Egl-GFP. The procedure for immunoprecipitation followed by RT-PCR was previously described (Sanghavi et al 2013). The RNA that coprecipitated with GFP and Egl-GFP was reverse transcribed using random hexamers and Superscript III (Life Technologies) using the directions provided by the manufacturer.…”

Section: Analyzing Mrna Associationmentioning

confidence: 99%

“…We have successfully used this approach to analyze the role of Dhc and the Dynein regulator, Dynamitin, in mRNA localization and endocytosis in midstage egg chambers (Sanghavi et al 2013;Liu et al 2015). A strain expressing an shRNA against egl (egl shRNA-1) was obtained from the Bloomington Drosophila Stock Center.…”

Section: Depletion Of Egl In Midstage Egg Chambersmentioning

confidence: 99%

“…This driver is present on the second chromosome, and is expressed at later stages of egg chamber maturation, from stages 5 or 6 onwards (Sanghavi et al 2013) (Figure S1B). For simplicity, we will refer to this strain as the "mid-stage driver."…”

Section: The Role Of Egl In Microtubule Organizationmentioning

confidence: 99%

“…Our results therefore suggest that, much like in the embryo, Egl functions in mRNA localization within the oocyte. Because the efficient localization of all three mRNAs requires the Dynein motor (Duncan and Warrior 2002;Januschke et al 2002;MacDougall et al 2003;Weil et al 2006;Sanghavi et al 2013), it is likely that Egl performs this function via its association with Dynein (Navarro et al 2004). …”

Section: Egl and Mrna Localizationmentioning

confidence: 99%

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Multiple Roles for Egalitarian in Polarization of the Drosophila Egg Chamber

et al. 2016

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The Drosophila egg chamber provides a useful model for examining mechanisms by which cell fates are specified and maintained in the context of a complex tissue. The egg chamber is also an excellent model for understanding the mechanism by which cytoskeletal filaments are organized and the critical interplay between cytoskeletal organization, polarity establishment, and cell fate specification. Previous work has shown that Egalitarian (Egl) is required for specification and maintenance of oocyte fate. Mutants in egl either completely fail to specify an oocyte, or if specified, the oocyte eventually reverts back to nurse cell fate. Due to this very early role for Egl in egg chamber maturation, it is unclear whether later stages of egg chamber development also require Egl function. In this report, we have depleted Egl at specific stages of egg chamber development. We demonstrate that in early-stage egg chambers, Egl has an additional role in organization of oocyte microtubules. In the absence of Egl function, oocyte microtubules completely fail to reorganize. As such, the localization of microtubule motors and their cargo is disrupted. In addition, Egl also appears to function in regulating the translation of critical polarity determining messenger RNAs (mRNAs). Finally, we demonstrate that in midstage egg chambers, Egl does not appear to be required for microtubule organization, but rather for the correct spatial localization of oskar, bicoid, and gurken mRNAs.

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“…In situ hybridization osk, bcd, and grk mRNAs were visualized as previously described (Sanghavi et al 2013).…”

Section: Immunofluorescencementioning

confidence: 99%

Section: Analyzing Mrna Associationmentioning

confidence: 99%

Section: Depletion Of Egl In Midstage Egg Chambersmentioning

confidence: 99%

Section: The Role Of Egl In Microtubule Organizationmentioning

confidence: 99%

Section: Egl and Mrna Localizationmentioning

confidence: 99%

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Multiple Roles for Egalitarian in Polarization of the Drosophila Egg Chamber

et al. 2016

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shot regulates the microtubule reorganization required for localization of axis‐determining mRNAs during oogenesis

et al. 2016

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The Drosophila mid-oogenesis stages are notable as the time when most maternal mRNAs become localized at discrete regions of the oocyte. Microtubule rearrangement occurs during this period and is critical for the localization of axis-determining maternal mRNAs. We have identified shot as a key player in establishing the cytoskeletal arrangement required for the spatial localization of axis-determining maternal mRNAs. We also found that the spatial distribution of the Shot protein is regulated by its mRNA localization. Our results suggest that the RNA localization mechanism is used not only for restricted accumulation of patterning molecules but also for the microtubule organization that leads to the initial development of oocyte polarity.

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The Role of Microtubule Motors in mRNA Localization and Patterning Within the Drosophila Oocyte

Goldman

Gonsalvez

2017

Results and Problems in Cell Differentiation

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Messenger RNA (mRNA) localization is a powerful and prevalent mechanism of post-transcriptional gene regulation, enabling the cell to produce protein at the exact location at which it is needed. The phenomenon of mRNA localization has been observed in many types of cells in organisms ranging from yeast to man. Thus, the process appears to be widespread and highly conserved. Several model systems have been used to understand the mechanism by which mRNAs are localized. One such model, and the focus of this chapter, is the egg chamber of the female Drosophila melanogaster. The polarity of the developing Drosophila oocyte and resulting embryo relies on the specific localization of three critical mRNAs: gurken, bicoid, and oskar. If these mRNAs are not localized during oogenesis, the resulting progeny will not survive. The study of these mRNAs has served as a model for understanding the general mechanisms by which mRNAs are sorted. In this chapter, we will discuss how the localization of these mRNAs enables polarity establishment. We will also discuss the role of motor proteins in the localization pathway. Finally, we will consider potential mechanisms by which mRNAs can be anchored at their site of localization. It is likely that the lessons learned using the Drosophila oocyte model system will be applicable to mRNAs that are localized in other organisms as well.

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Dynein Associates with oskar mRNPs and Is Required For Their Efficient Net Plus-End Localization in Drosophila Oocytes

Cited by 26 publications

References 56 publications

Multiple Roles for Egalitarian in Polarization of the Drosophila Egg Chamber

Multiple Roles for Egalitarian in Polarization of the Drosophila Egg Chamber

shot regulates the microtubule reorganization required for localization of axis‐determining mRNAs during oogenesis

The Role of Microtubule Motors in mRNA Localization and Patterning Within the Drosophila Oocyte

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