Staufen2 mediated RNA recognition and localization requires combinatorial action of multiple domains

Heber, Simone; Gáspár, Imre; Tants, Jan‐Niklas; Günther, Johannes; Moya, Sandra M. Fernandez; Janowski, Robert; Ephrussi, Anne; Sattler, Michael; Niessing, Dierk

doi:10.1101/396994

Cited by 3 publications

(9 citation statements)

References 52 publications

(53 reference statements)

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“…To test if a regulatory circuit built around Staufen and Egalitarian also acts on other transcripts, we analyzed the localization and composition of the anterior localizing bicoid mRNPs (Fig 6A; Berleth et al, 1988), a known target of Staufen (Ferrandon et al, 1994) and a likely target of Egalitarian (Dienstbier et al, 2009;Ferrandon et al, 1994) . As previously reported, bicoid mRNPs may also contain multiple mRNA copies (Ferrandon et al, 1994;Heber et al, 2019;Trovisco et al, 2016 and Fig 6B and S5B), similar to oskar mRNPs. We found that the relative amount of Staufen-associated bicoid mRNPs is largely independent of RNA copy number (Fig 6B).…”

Section: Localization Of Bicoid Mrna Is Regulated By Staufen and Egalitariansupporting

confidence: 84%

“…Quantifying oskar mRNA copy number by smFISH (Fig S3A ), we found that the fraction of Staufen-associated oskar mRNPs in the oocyte increased as a function of oskar mRNA content, and the increase was substantially more pronounced during stage 9 than in earlier stages of oogenesis (Fig 3B). During stage 9, not only the likelihood of Staufen association but also the relative levels of Staufen -as reported by the normalised Staufen-GFP signal -increased and this increase was highly linear (Fig 3B' and Fig S3C), similarly to what we reported for overexpressed Staufen and mammalian Stau2 (Heber et al, 2019). Prior to the onset of oskar localization, Staufen levels showed no scaling with oskar mRNA content of the mRNPs (Fig 3B').…”

Section: Staufen Dissociates Egalitarian From Oskar Mrnps In the Oocytesupporting

confidence: 81%

“…Analysis was carried out as described in (Gáspár et al, 2017;Heber et al, 2019). Briefly, images for colocalization analysis were deconvolved using Huygens Essential and segmented using a custom ImageJ plug-in.…”

Section: Colocalization Analysis Between the Mrnps And Fluorescent Reportersmentioning

confidence: 99%

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An RNA-based feed-forward mechanism ensures motor switching in oskar mRNA transport

Gáspár

Ephrussi

2021

Preprint

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The regulated recruitment and activity of motor proteins is crucial for intracellular transport of cargoes, including ribonucleoprotein complexes (RNP). Here we show that regulation of mRNP transport by the minus end-directed dynein motor complex in the Drosophila germline relies on the interplay of two double-stranded RNA binding proteins, Staufen and the dynein adaptor Egalitarian. Our quantitative in situ analysis shows that, in the nurse cells, Egalitarian associates with bicoid, oskar and staufen mRNAs, which consequently enrich in the oocyte. This results in ooplasmic accumulation of Staufen and its recruitment to the transcripts. We demonstrate that Staufen levels scale proportionately with RNA content in oskar mRNPs, and to a lesser extent in bicoid mRNPs. We show that enhanced recruitment of Staufen to oskar mRNPs results in dissociation of Egalitarian and a reduction in minus end-directed transport, although dynein remains associated with the RNPs. The downregulation of dynein activity prevents anterior accumulation of oskar mRNA, whose transport to the posterior is essential for localized production of Oskar protein. Our observations identify a feed-forward loop, whereby staufen mRNA localization and protein accumulation in the oocyte enable motor switching, promoting oskar mRNA localization to the posterior pole.

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Section: Localization Of Bicoid Mrna Is Regulated By Staufen and Egalitariansupporting

confidence: 84%

Section: Staufen Dissociates Egalitarian From Oskar Mrnps In the Oocytesupporting

confidence: 81%

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An RNA-based feed-forward mechanism ensures motor switching in oskar mRNA transport

Gáspár

Ephrussi

2021

Preprint

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“…Notably, BD3 and RBD4 regulate STAU1 RNA-binding activity (Wickham et al, 1999) and RBD4-TBD is involved in ribosome association (Luo et al, 2002). We now show that STAU1 association with spindles requires the N-terminal region that contains RBD2, a domain devoid of RNA-binding activity in vitro (Wickham et al, 1999), although we do not exclude the possibility that RBD2 could bind RNA in vivo as reported for the paralogue protein STAU2 (Heber et al, 2019). This result indicates that STAU1 RNAbinding and ribosome-binding activities are not involved in spindle association.…”

Section: Discussionmentioning

confidence: 60%

Staufen1 localizes to the mitotic spindle and controls the localization of RNA populations to the spindle

Hassine

Bonnet-Magnaval

Bouvrette

et al. 2020

Journal of Cell Science

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Staufen1 (STAU1) is an RNA-binding protein involved in the post-transcriptional regulation of mRNAs. We report that a large fraction of STAU1 localizes to the mitotic spindle in colorectal cancer HCT116 cells and in non-transformed hTERT-RPE1 cells. Spindle-associated STAU1 partly co-localizes with ribosomes and active sites of translation. We mapped the molecular determinant required for STAU1–spindle association within the first 88 N-terminal amino acids, a domain that is not required for RNA binding. Interestingly, transcriptomic analysis of purified mitotic spindles revealed that 1054 mRNAs and the precursor ribosomal RNA (pre-rRNA), as well as the long non-coding RNAs and small nucleolar RNAs involved in ribonucleoprotein assembly and processing, are enriched on spindles compared with cell extracts. STAU1 knockout causes displacement of the pre-rRNA and of 154 mRNAs coding for proteins involved in actin cytoskeleton organization and cell growth, highlighting a role for STAU1 in mRNA trafficking to spindle. These data demonstrate that STAU1 controls the localization of subpopulations of RNAs during mitosis and suggests a novel role of STAU1 in pre-rRNA maintenance during mitosis, ribogenesis and/or nucleoli reassembly.

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“…Koh et al have shown involvement of dynamics at the dsRNA-dsRBD interface where the diffusion of dsRBDs of TRBP along pre-miRNA favor its cleavage to the miRNA:miRNA* duplex by Dicer -an RNase III enzyme (Koh et al, 2013). Heber et al have recently compared binding affinities and binding stabilities of various dsRBDs in a multi-domain protein for RNA-recognition in Drosophila Staufen protein (Heber et al, 2019). Fareh et al have shown that the recognition of target dsRNA by TRBP involves dual binding modes to discriminate between the target dsRNA and other cellular dsRNAs to which protein is exposed (Fareh et al, 2016).…”

Section: Introductionmentioning

confidence: 99%

Conformational dynamics at microsecond timescale in the RNA-binding regions of dsRNA-binding domains

Paithankar¹,

Chugh²

2019

Preprint

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Double-stranded RNA-binding domains (dsRBDs) are involved in a variety of biological functions via recognition and processing of dsRNAs. Though the primary substrate of the dsRBDs are dsRNAs with A-form helical geometry; they are known to interact with structurally diverse dsRNAs. Here, we have employed two model dsRBDs -TAR-RNA binding protein and Adenosine deaminase that acts on RNA -to understand the role of intrinsic protein dynamics in RNA binding. We have performed a detailed characterization of the residue-level dynamics by NMR spectroscopy for the two dsRBDs.While the dynamics profiles at the ps-ns timescale of the two dsRBDs were found to be different, a striking similarity was observed in the μ s-ms timescale dynamics for both the dsRBDs. Motions at fast μ s timescale (k ex > 50000 s -1 ) were found to be present not only in the RNA-binding residues but also in some allosteric residues of the dsRBDs. We propose that this intrinsic μ s timescale dynamics in two distinct dsRBDs allows them to undergo conformational rearrangement that may aid dsRBDs to target substrate dsRNA from the pool of structurally different RNAs in cellular environment.

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Staufen2 mediated RNA recognition and localization requires combinatorial action of multiple domains

Cited by 3 publications

References 52 publications

An RNA-based feed-forward mechanism ensures motor switching in oskar mRNA transport

An RNA-based feed-forward mechanism ensures motor switching in oskar mRNA transport

Staufen1 localizes to the mitotic spindle and controls the localization of RNA populations to the spindle

Conformational dynamics at microsecond timescale in the RNA-binding regions of dsRNA-binding domains

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