Egalitarian binding partners, Dynein Light Chain and Bicaudal-D, act sequentially to link mRNA to the Dynein motor

Goldman, Chandler H.; Veeranan-Karmegam, Rajalakshmi; Neiswender, Hannah; Gonsalvez, Graydon B.

doi:10.1101/550277

Cited by 6 publications

(24 citation statements)

References 39 publications

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“…In support of this model, we showed that artificially dimerizing a mutant version of Egl that could not associate with Dlc was sufficient to restore its ability to bind to mRNA and BicD. These experiments were conducted using lysates from Drosophila S2 cells (Goldman et al, 2019). An important question raised by this finding is whether artificially dimerizing Egl could rescue mutant phenotypes in vivo.…”

Section: Introductionmentioning

confidence: 84%

“…The in vitro RNA binding experiments using ILS, TLS and GLS localization elements were performed as previously described (Dienstbier et al, 2009;Goldman et al, 2019).…”

Section: Rna Binding Studiesmentioning

confidence: 99%

“…Mutations in Egl that compromised Dlc binding were defective for dimerization. These mutants also displayed reduced RNA binding activity, and as a consequence, BicD binding was reduced (Goldman et al, 2019).…”

Section: Introductionmentioning

confidence: 99%

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Dynein light chain-dependent dimerization of Egalitarian is essential for maintaining oocyte fate in Drosophila

Neiswender

Goldman

Veeranan-Karmegam

et al. 2021

Preprint

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Egalitarian (Egl) is an RNA adaptor for the Dynein motor and is thought to link numerous, perhaps hundreds, of mRNAs with Dynein. Dynein, in turn, is responsible for the transport and localization of these mRNAs. Studies have shown that efficient mRNA binding by Egl requires the protein to dimerize. We recently demonstrated that Dynein light chain (Dlc) is responsible for facilitating the dimerization of Egl. Mutations in Egl that fail to interact with Dlc do not dimerize, and as such, are defective for mRNA binding. Consequently, this mutant does not efficiently associate with BicaudalD (BicD), the factor responsible for linking the Egl/mRNA complex with Dynein. In this report, we tested whether artificially dimerizing this Dlc-binding mutant using a leucine zipper would restore mRNA binding and rescue mutant phenotypes in vivo. Interestingly, we found that although artificial dimerization of Egl restored BicD binding, it only partially restored mRNA binding. As a result, Egl-dependent phenotypes, such as oocyte specification and mRNA localization, were only partially rescued. We hypothesize that Dlc-mediated dimerization of Egl results in a three-dimensional conformation of the Egl dimer that is best suited for mRNA binding. Although the leucine zipper restores Egl dimerization, it likely does not enable Egl to assemble into the conformation required for maximal mRNA binding activity.

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Section: Introductionmentioning

confidence: 84%

“…The in vitro RNA binding experiments using ILS, TLS and GLS localization elements were performed as previously described (Dienstbier et al, 2009;Goldman et al, 2019).…”

Section: Rna Binding Studiesmentioning

confidence: 99%

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Dynein light chain-dependent dimerization of Egalitarian is essential for maintaining oocyte fate in Drosophila

Neiswender

Goldman

Veeranan-Karmegam

et al. 2021

Preprint

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“…Ovaries were processed for in situ hybridization using a published protocol (Goldman et al, 2019). Dissected ovaries were fixed with 4% formaldehyde for 20 mins.…”

Section: In Situ Hybridizationmentioning

confidence: 99%

“…Ovaries were processed for immunofluorescence using a previous described procedure (Goldman et al, 2019). Briefly, flies were fattened on yeast pellets for 3 days.…”

Section: Immunofluorescencementioning

confidence: 99%

A region of Drosophila SLBP distinct from the histone pre-mRNA binding and processing domains is essential for deposition of histone mRNA in the oocyte

Potter-Birriel¹

2020

Preprint

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During Drosophila oogenesis, large amounts of histone mRNA and proteins are deposited in the developing oocyte. These are sufficient for the first 14 embryonic cell cycles and provide the developing embryo with sufficient histone proteins until the zygotic histone genes are activated. The maternally deposited histone mRNA is synthesized in stage 10b of oogenesis after completion of endoreduplication of the nurse cells. Histone mRNAs are the only cellular mRNAs that are not polyadenylated, ending instead in a conserved stemloop instead of a polyA tail. The Stem-loop binding protein (SLBP) binds the 3' end of histone mRNA and is essential for both the biosynthesis and translation of histone mRNA. We report that a 10 aa region in SLBP, which is not required for processing in vitro, is essential for transcription of histone mRNA in the stage 10b oocyte. In stage 10b the Histone Locus Bodies (HLBs) produce histone mRNAs in the absence of phosphorylation of Mxc, normally required for histone gene expression in S-phase cells. Mutants expressing this SLBP develop normally, produce small amounts of polyadenylated histone mRNA throughout development, but little histone mRNA in stage 10b resulting in death of the embryos in the first hr of development.

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Patterning the Drosophila embryo: A paradigm for RNA‐based developmental genetic regulation

Lasko

2020

WIREs RNA

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Embryonic anterior-posterior patterning is established in Drosophila melanogaster by maternally expressed genes. The mRNAs of several of these genes accumulate at either the anterior or posterior pole of the oocyte via a number of mechanisms. Many of these mRNAs are also under elaborate translational regulation. Asymmetric RNA localization coupled with spatially restricted translation ensures that their proteins are restricted to the position necessary for the developmental process that they drive. Bicoid (Bcd), the anterior determinant, and Oskar (Osk), the determinant for primordial germ cells and posterior patterning, have been studied particularly closely. In early embryos an anterior-posterior gradient of Bcd is established, activating transcription of different sets of zygotic genes depending on local Bcd concentration. At the posterior pole, Osk seeds formation of polar granules, ribonucleoprotein complexes that accumulate further mRNAs and proteins involved in posterior patterning and germ cell specification. After fertilization, polar granules associate with posterior nuclei and mature into nuclear germ granules. Osk accumulates in these granules, and either by itself or as part of the granules, stimulates germ cell division.

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Egalitarian binding partners, Dynein Light Chain and Bicaudal-D, act sequentially to link mRNA to the Dynein motor

Cited by 6 publications

References 39 publications

Dynein light chain-dependent dimerization of Egalitarian is essential for maintaining oocyte fate in Drosophila

Dynein light chain-dependent dimerization of Egalitarian is essential for maintaining oocyte fate in Drosophila

A region of Drosophila SLBP distinct from the histone pre-mRNA binding and processing domains is essential for deposition of histone mRNA in the oocyte

Patterning the Drosophila embryo: A paradigm for RNA‐based developmental genetic regulation

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