Crystal structure of zinc-finger domain of Nanos and its functional implications

Hashimoto, H.; Hara, K.; Hishiki, Asami; Kawaguchi, S.; Yoshizawa, T.; Unzai, Satoru; Tamaru, Yutaka; Shimizu, Takayuki; Sato, Mitsuaki

doi:10.1107/s0108767311094566

Cited by 5 publications

(5 citation statements)

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“…However, the isolated NED did not repress the expression of this reporter (Fig E and F), most likely because it lacks RNA binding capacity. These results indicate that the Nanos NED confers repressive activity but requires the ZnF domain to bind to natural mRNA targets, as reported previously (Curtis et al , ; Hashimoto et al , ).…”

Section: Resultssupporting

confidence: 89%

“…Three Nanos paralogs (Nanos1-3) exist in vertebrates and various invertebrate species, whereas there is only one family member in Dm and other insects (Subramaniam & Seydoux, 1999;Mochizuki et al, 2000;Jaruzelska et al, 2003;Tsuda et al, 2003). This protein family is defined by a highly conserved CCHC-type zinc-finger (ZnF) domain (Curtis et al, 1997;Hashimoto et al, 2010) and divergent N-and C-terminal unstructured regions of variable lengths ( Fig 1A). The ZnF domain mediates binding to RNA and to Pumilio, a conserved Nanos partner that confers mRNA target specificity (Murata & Wharton, 1995;Curtis et al, 1997;Wreden et al, 1997;Asaoka-Taguchi et al, 1999;Sonoda & Wharton, 1999;Jaruzelska et al, 2003).…”

Section: Introductionmentioning

confidence: 99%

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Distinct modes of recruitment of the CCR 4– NOT complex by Drosophila and vertebrate Nanos

et al. 2016

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Nanos proteins repress the expression of target mRNAs by recruiting effector complexes through non‐conserved N‐terminal regions. In vertebrates, Nanos proteins interact with the NOT1 subunit of the CCR4–NOT effector complex through a NOT1 interacting motif (NIM), which is absent in Nanos orthologs from several invertebrate species. Therefore, it has remained unclear whether the Nanos repressive mechanism is conserved and whether it also involves direct interactions with the CCR4–NOT deadenylase complex in invertebrates. Here, we identify an effector domain (NED) that is necessary for the Drosophila melanogaster (Dm) Nanos to repress mRNA targets. The NED recruits the CCR4–NOT complex through multiple and redundant binding sites, including a central region that interacts with the NOT module, which comprises the C‐terminal domains of NOT1–3. The crystal structure of the NED central region bound to the NOT module reveals an unanticipated bipartite binding interface that contacts NOT1 and NOT3 and is distinct from the NIM of vertebrate Nanos. Thus, despite the absence of sequence conservation, the N‐terminal regions of Nanos proteins recruit CCR4–NOT to assemble analogous repressive complexes.

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Section: Resultssupporting

confidence: 89%

Section: Introductionmentioning

confidence: 99%

Distinct modes of recruitment of the CCR 4– NOT complex by Drosophila and vertebrate Nanos

et al. 2016

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“…Most recently, the structure of the zincfinger region has been solved and shown to be required for RNA binding (Hashimoto et al, 2010). Together with Pumilio, Nanos is part of a translational repression complex in which Pumilio provides the RNA binding specificity and Nanos the repressive activity (Jaruzelska et al, 2003;Lai et al, 2011;Sonoda and Wharton, 1999;Sonoda and Wharton, 2001).…”

Section: Introductionmentioning

confidence: 99%

XenopusNanos1 is required to prevent endoderm gene expression and apoptosis in primordial germ cells

2012

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Nanos is expressed in multipotent cells, stem cells and primordial germ cells (PGCs) of organisms as diverse as jellyfish and humans. It functions together with Pumilio to translationally repress targeted mRNAs. Here we show by loss-of-function experiments that Xenopus Nanos1 is required to preserve PGC fate. Morpholino knockdown of maternal Nanos1 resulted in a striking decrease in PGCs and a loss of germ cells from the gonads. Lineage tracing and TUNEL staining reveal that Nanos1-deficient PGCs fail to migrate out of the endoderm. They appear to undergo apoptosis rather than convert to normal endoderm. Whereas normal PGCs do not become transcriptionally active until neurula, Nanos1-depleted PGCs prematurely exhibit a hyperphosphorylated RNA polymerase II C-terminal domain at the midblastula transition. Furthermore, they inappropriately express somatic genes characteristic of endoderm regulated by maternal VegT, including Xsox17α, Bix4, Mixer, GATA4 and Edd. We further demonstrate that Pumilio specifically binds VegT RNA in vitro and represses, along with Nanos1, VegT translation within PGCs. Repressed VegT RNA in wild-type PGCs is significantly less stable than VegT in Nanos1-depleted PGCs. Our data indicate that maternal VegT RNA is an authentic target of Nanos1/Pumilio translational repression. We propose that Nanos1 functions to translationally repress RNAs that normally specify endoderm and promote apoptosis, thus preserving the germline.

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“…An alignment of the echinoderm Nanos protein sequences reveal each protein has two conserved CCHC zinc fingers that are characteristic of the Nanos protein family and are necessary for function (Fig. A, asterisks) (Arrizabalaga and Lehmann, ; Hashimoto et al, ). In addition, echinoderm Nanos proteins contain a conserved upstream domain that is found in non‐protostome Nanos sequences and is also required for function (Fig.…”

Section: Resultsmentioning

confidence: 99%

The diversity of nanos expression in echinoderm embryos supports different mechanisms in germ cell specification

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Swartz

Juliano

et al. 2016

Evolution and Development

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Specification of the germ cell lineage is required for sexual reproduction in all animals. However, the timing and mechanisms of germ cell specification is remarkably diverse in animal development. Echinoderms, such as sea urchins and sea stars, are excellent model systems to study the molecular and cellular mechanisms that contribute to germ cell specification. In several echinoderm embryos tested, the germ cell factor Vasa accumulates broadly during early development and is restricted after gastrulation to cells that contribute to the germ cell lineage. In the sea urchin, however, the germ cell factor Vasa is restricted to a specific lineage by the 32-cell stage. We therefore hypothesized that the germ cell specification program in the sea urchin/Euechinoid lineage has evolved to an earlier developmental time point. To test this hypothesis we determined the expression pattern of a second germ cell factor, Nanos, in four out of five extant echinoderm clades. Here we find that Nanos mRNA does not accumulate until the blastula stage or later during the development of all other echinoderm embryos except those that belong to the Echinoid lineage. Instead, Nanos is expressed in a restricted domain at the 32–128 cell stage in Echinoid embryos. Our results support the model that the germ cell specification program underwent a heterochronic shift in the Echinoid lineage. A comparison of Echinoid and non-Echinoid germ cell specification mechanisms will contribute to our understanding of how these mechanisms have changed during animal evolution.

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Crystal structure of zinc-finger domain of Nanos and its functional implications

Cited by 5 publications

References 0 publications

Distinct modes of recruitment of the CCR 4– NOT complex by Drosophila and vertebrate Nanos

Distinct modes of recruitment of the CCR 4– NOT complex by Drosophila and vertebrate Nanos

XenopusNanos1 is required to prevent endoderm gene expression and apoptosis in primordial germ cells

The diversity of nanos expression in echinoderm embryos supports different mechanisms in germ cell specification

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