Functional dissection of the RNA signal sequence responsible for vegetal localization of<i>XGrip2.1</i>mRNA in Xenopus oocytes

Claußen, Maike; Tarbashevich, Katsiaryna; Pieler, Tomas

doi:10.4161/rna.8.5.16028

Cited by 13 publications

(9 citation statements)

References 40 publications

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“…Conversely, no binding was detected with two other LE RNAs derived from either velo1 or gdf1. Ptbp1 served as control as it has been shown to interact with the dnd1-LE, the grip2-LE and the gdf1-LE (Claussen et al, 2011).…”

Section: Celf1 Binds To Specific Rna-les In Vitro and It Is In A Compmentioning

confidence: 99%

“…4A, B) were used. 0.4 pmol Cy3-labeled RNA was incubated with different concentrations of recombinant proteins in a 20 ml reaction mix containing 1x UV-crosslinking-buffer (Claussen et al, 2011) for 30 min at RT. 5 ml loading buffer (50% glycerol, 50 mM Tris/HCl pH 7.5, 0.01% bromphenol blue) was added and the whole volume was loaded on a 1% agarose Tris/ acetic acid/EDTA buffer gel.…”

Section: Electrophoretic Mobility Shift Assays (Emsas)mentioning

confidence: 99%

“…Precipitated proteins were separated by SDS-PAGE and visualized by Western blot. Co-immunoprecipitation of Cy3-labeled RNAs was done as described in Claussen et al (2011).…”

Section: Co-immunoprecipitationsmentioning

confidence: 99%

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A novel role for Celf1 in vegetal RNA localization during Xenopus oogenesis

Bauermeister

Claußen

Pieler

2015

Developmental Biology

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The localization of certain mRNAs to the vegetal cortex of Xenopus oocytes is of crucial importance for germ cell development and early embryonic patterning. Vegetal RNA localization is mediated by cis-acting RNA localization elements (LE). Several proteins assemble on the RNA LE and direct transport to the vegetal cortex. Although a number of localization RNP components have been identified, their full composition is unknown. In an RNA affinity purification approach, using the dead end 1 (dnd1) RNA LE, we identified Xenopus Celf1 as a novel component of vegetal localization RNP complexes. Celf1 is part of an RNP complex together with known vegetal localization factors and shows specific interactions with LEs from several but not all vegetally localizing RNAs. Immunostaining experiments reveal co-localization of Celf1 with vegetally localizing RNA and with known localization factors. Inhibition of Celf1 protein binding by localization element mutagenesis as well as Celf1 overexpression interfere with vegetal RNA localization. These results argue for a role of Celf1 in vegetal RNA localization during Xenopus oogenesis.

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Section: Celf1 Binds To Specific Rna-les In Vitro and It Is In A Compmentioning

confidence: 99%

Section: Electrophoretic Mobility Shift Assays (Emsas)mentioning

confidence: 99%

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A novel role for Celf1 in vegetal RNA localization during Xenopus oogenesis

Bauermeister

Claußen

Pieler

2015

Developmental Biology

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“…RNAs highly concentrated at the vegetal pole were found enriched in an oocyte cytokeratin fraction and led to the first molecular cloning of Xenopus germ plasm RNAs (Houston et al 1998;MacArthur et al 2000;Mosquera et al 1993;Pondel and King 1988;Zhang and King 1996). More recently, microarray analysis of RNA from isolated vegetal cortices has yielded over a hundred candidates (Claussen et al 2011;Cuykendall and Houston 2010). Some of these have now been confirmed by in situ hybridization as being specifically localized in the MC and later in oocyte germ plasm islands or embryonic germ plasm.…”

Section: Molecular Components Of Germ Plasmmentioning

confidence: 97%

Germ‐Cell Specification inXenopus

King

2014

Xenopus Development

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Primordial germ cells (PGCs) represent the only precursor stem cell population that normally gives rise to the gametes. For this reason, PGCs have been called the "stem cells of the species" and are prime examples of a normal totipotent state (Wylie, 1999. Cell 96 (2): 165-174). PGCs represent a remarkable lineage that will differentiate into highly specialized cells, either oocytes or sperm, yet also retain the ability to generate a complete new individual. Thus, a fundamental question in developmental and stem cell biology is how PGCs differentiate while retaining the potential for totipotency in contrast to somatic cells that become fate-restricted. In this chapter, we will review what is known about how Xenopus sets aside its germ-cell lineage.

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“…How are PGC mRNAs controlled at the translation level? A recurrent theme is how common vegetally localized mRNAs also play a role in neurogenic fates or functions (Owens et al, ). In fact, the most abundant mRNA within the vegetal cortex is grip2 (glutamate receptor interacting protein), a scaffold multiple PDZ domain protein that organizes signaling complexes in neurons (Ge et al, ; Kaneshiro, Miyauchi, Tanigawa, Ikenishi, & Komiya, ; Claussen et al, ; Tan, Queenan, & Huganir, ; Tarbashevich, Koebernick, & Pieler, ; Kirilenko et al, ). Grip2 protein is present within vegetal cortices and may well serve as a scaffold for signaling at the vegetal pole providing the link between Wnt and PGC migration (unpublished observations, Newman & King).…”

Section: Future Directionsmentioning

confidence: 99%

Maternal messages to live by: a personal historical perspective

King

2017

Genesis

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SUMMARY In the 1980’s, the study of localized maternal mRNAs was just emerging as a new research area. Classic embryological studies had linked the inheritance of cytoplasmic domains with specific cell lineages, but the underlying molecular nature of these putative determinants remained a mystery. The model system Xenopus would play a pivotal role in the progress of this new field. In fact, the first localized maternal mRNA to be identified and cloned from any organism was Xenopus vg1, a TGF-beta family member. This seminal finding opened the door to many subsequent studies focused on how RNAs are localized and what functions they had in development. Xenopus remains the model of choice for studies on dissecting RNA/protein transport particles and RNA-sequencing analyses that are moving the field into the future.

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Functional dissection of the RNA signal sequence responsible for vegetal localization ofXGrip2.1mRNA in Xenopus oocytes

Cited by 13 publications

References 40 publications

A novel role for Celf1 in vegetal RNA localization during Xenopus oogenesis

A novel role for Celf1 in vegetal RNA localization during Xenopus oogenesis

Germ‐Cell Specification inXenopus

Maternal messages to live by: a personal historical perspective

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