The nanos translational control element represses translation in somatic cells by a Bearded box-like motif

Duchow, Heather K.; Brechbiel, Jillian L.; Chatterjee, Seema; Gavis, Elizabeth R.

doi:10.1016/j.ydbio.2005.03.025

Cited by 13 publications

(12 citation statements)

References 39 publications

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“…Double fluorescence in situ hybridization (FISH) using tyramide signal amplification was performed as described (Kosman et al, 2004). Immunofluorescence was performed as previously described (Duchow et al, 2005) using embryos heat-fixed in 68 mM NaCl/0.03% Triton X-100, with rabbit anti-Vas (1:10000; gift from R. Lehmann) and Alexa Fluor-568 goat anti-rabbit (1:1000; Molecular Probes). Embryos were mounted in 90% glycerol/100 mM Tris pH 8.0 and imaged with a Zeiss LSM510 confocal microscope.…”

Section: Methodsmentioning

confidence: 99%

Bazooka regulates microtubule organization and spatial restriction of germ plasm assembly in the Drosophila oocyte

Becalska

Gavis

2010

Developmental Biology

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Localization of the germ plasm to the posterior of the Drosophila oocyte is required for anteroposterior patterning and germ cell development during embryogenesis. While mechanisms governing the localization of individual germ plasm components have been elucidated, the process by which germ plasm assembly is restricted to the posterior pole is poorly understood. In this study, we identify a novel allele of baz, the Drosophila homolog of Par-3, that has allowed the analysis of baz function throughout oogenesis. We demonstrate that baz is required for spatial restriction of the germ plasm and axis patterning and we uncover multiple requirements for baz in regulating the organization of the oocyte microtubule cytoskeleton. Our results suggest that distinct cortical domains established by Par proteins polarize the oocyte through differential effects on microtubule organization. We further show that microtubule plus-end enrichment is sufficient to drive germ plasm assembly even at a distance from the oocyte cortex, suggesting that control of microtubule organization is critical not only for the localization of germ plasm components to the posterior of the oocyte but also for the restriction of germ plasm assembly to the posterior pole.

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Section: Methodsmentioning

confidence: 99%

Bazooka regulates microtubule organization and spatial restriction of germ plasm assembly in the Drosophila oocyte

Becalska

Gavis

2010

Developmental Biology

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“…We expect that excess Nanos2 protein may negatively affect its own translational efficiency, but we have shown that the Nanos2-3Ј-UTR might be required for efficient translation also (Tsuda et al, 2006). Because the 3ЈUTR elements within the Nanos family of mRNAs have also been shown to be regulated by several proteins (Dahanukar and Wharton, 1996;de Moor et al, 2005;Duchow et al, 2005;Nelson et al, 2004), these factors may also affect the translational efficiency of Nanos2 in combination with other mechanisms.…”

Section: Research Articlementioning

confidence: 97%

Functional redundancy among Nanos proteins and a distinct role of Nanos2 during male germ cell development

2007

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The mouse Nanos proteins, Nanos2 and Nanos3, are required for germ cell development and share a highly conserved zinc-finger domain. The expression patterns of these factors during development, however, differ from each other. Nanos3 expression in the mouse embryo commences in the primordial germ cells (PGCs) just after their formation, and a loss of this protein results in the germ cell-less phenotype in both sexes. By contrast, Nanos2 expression begins only in male PGCs after their entry into the genital ridge and a loss of this protein results in a male germ cell deficiency, irrespective of the co-expression of Nanos3 in these cells. These results indicate that these two Nanos proteins have distinct functions, which depend on the time and place of their expression. To further elucidate this, we have generated transgenic mouse lines that express Nanos2 under the control of the Oct4⌬PE promoter and examined Nanos2 function in a Nanos3-null genetic background. We find that ectopically produced Nanos2 protein rescues the Nanos3-null defects, because the germ cells fully develop in both sexes in the transgenic mice. This result indicates that Nanos2 can substitute for Nanos3 during early PGC development. By contrast, our current data show that Nanos3 does not rescue the defects in Nanos2-null mice. Our present findings thus indicate that there are redundant functions of the Nanos proteins in early PGC development, but that Nanos2 has a distinct function during male germ cell development in the mouse.

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“…Immunostaining of ovaries was performed as previously described (Shcherbata et al, 2004). Embryo immunostaining was performed as described (Duchow et al, 2005), except that for anti-Rump immunofluorescence, dechorionated embryos were heat-fixed in 68 mM NaCl/0.03% Triton X-100, transferred to PBS, and the vitelline membranes were manually removed using a 30-gauge needle. The following antibodies and dyes were used: 1:100 anti-Rump 10C3 (immunofluorescence), 1:100 anti-Rump 5G4 (immunohistochemistry), 1:10,000 rabbit anti-Vas (gift of R. Lehmann, Skirball Institute, New York), 1:1000 Alexa Fluor 568 goat anti-mouse (Invitrogen/Molecular Probes), 1:1000 Alexa Fluor 488 goat anti-rabbit (Invitrogen/Molecular Probes), 1:1000 Oregon Green 488 phalloidin (Invitrogen/Molecular Probes), 1:1000 Hoechst (ovaries), 1:1000 DAPI (embryos).…”

Section: Embryonic Cuticle Preparation In Situ Hybridization and Immmentioning

confidence: 99%

TheDrosophilahnRNP M homolog Rumpelstiltskin regulatesnanosmRNA localization

Jain

Gavis

2008

Development

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Anterior-posterior axis patterning of the Drosophila embryo requires Nanos activity selectively in the posterior. This spatial asymmetry of Nanos is generated by the localization of nanos mRNA to the posterior pole of the embryo, where it is subsequently translated. Posterior localization of nanos is mediated by a complex cis-acting localization signal in its 3Ј untranslated region comprising several partially redundant localization elements. This localization signal redundancy has hampered the identification of trans-acting factors that act specifically to effect posterior localization of nanos. Here, we have used a biochemical approach to identify Rumpelstiltskin, a Drosophila heterogeneous nuclear ribonucleoprotein (hnRNP) M homolog, which binds directly to an individual nanos localization element. Rumpelstiltskin associates with nanos mRNA in vitro and in vivo, and binding by Rumpelstiltskin correlates with localization element function in vivo. Through analysis of a rumpelstiltskin null mutation by genetic strategies that circumvent redundancy, we demonstrate that Rumpelstiltskin regulates anterior-posterior axis patterning by functioning as a direct-acting nanos mRNA localization factor.

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The nanos translational control element represses translation in somatic cells by a Bearded box-like motif

Cited by 13 publications

References 39 publications

Bazooka regulates microtubule organization and spatial restriction of germ plasm assembly in the Drosophila oocyte

Bazooka regulates microtubule organization and spatial restriction of germ plasm assembly in the Drosophila oocyte

Functional redundancy among Nanos proteins and a distinct role of Nanos2 during male germ cell development

TheDrosophilahnRNP M homolog Rumpelstiltskin regulatesnanosmRNA localization

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