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The Nanos3-3'UTR is required for germ cell specific NANOS3 expression in mouse embryos.

Suzuki H, Saba R, Sada A, Saga Y - PLoS ONE (2010)

Bottom Line: This is fundamental to the continuation of a species.Although Nanos3 is transcribed in both cell lineages, it is efficiently translated only in the germ lineage.Surprisingly, even when under the control of the CAG promoter which induces strong ubiquitous transcription in both germ cells and somatic cells, the addition of the Nanos3-3'UTR sequence to the coding region of exogenous gene was effective in restricting protein expression in germ cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, Graduate School of Science, University of Tokyo, Tokyo, Japan.

ABSTRACT

Background: The regulation of gene expression via a 3' untranslated region (UTR) plays essential roles in the discrimination of the germ cell lineage from somatic cells during embryogenesis. This is fundamental to the continuation of a species. Mouse NANOS3 is an essential protein required for the germ cell maintenance and is specifically expressed in these cells. However, the regulatory mechanisms that restrict the expression of this gene in the germ cells is largely unknown at present.

Methodology/principal findings: In our current study, we show that differences in the stability of Nanos3 mRNA between germ cells and somatic cells is brought about in a 3'UTR-dependent manner in mouse embryos. Although Nanos3 is transcribed in both cell lineages, it is efficiently translated only in the germ lineage. We also find that the translational suppression of NANOS3 in somatic cells is caused by a 3'UTR-mediated mRNA destabilizing mechanism. Surprisingly, even when under the control of the CAG promoter which induces strong ubiquitous transcription in both germ cells and somatic cells, the addition of the Nanos3-3'UTR sequence to the coding region of exogenous gene was effective in restricting protein expression in germ cells.

Conclusions/significance: Our current study thus suggests that Nanos3-3'UTR has an essential role in translational control in the mouse embryo.

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Related in: MedlinePlus

The accumulation of somatic Nanos3 mRNA is suppressed by the Nanos3-3′UTR.The levels of Nanos3 (A–C) or mRFP (D) mRNA were compared by quantitative RT-PCR using RNA samples derived from wild-type embryos (A–C) and BAC-Nanos3-mRFP(Nos3-3′UTR) (blue in D) and BAC-Nanos3-mRFP(BghpA) (green in D) transgenic embryos at E7.5, E9.5 and E13.5. Data were normalized by G3PDH in each sample. The relative mRNA levels in the E9.5A sample (an anterior part of E9.5 embryo) were assigned the reference value of 1.0. A, anterior part of the embryo; P, posterior part of the embryo; M–K or K, the male kidney; M–G or G, the male gonad; F–G, the female gonad; N, the posterior part of a Nanos3 knockout embryo at E9.5. Error bars represent the s.d.
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pone-0009300-g004: The accumulation of somatic Nanos3 mRNA is suppressed by the Nanos3-3′UTR.The levels of Nanos3 (A–C) or mRFP (D) mRNA were compared by quantitative RT-PCR using RNA samples derived from wild-type embryos (A–C) and BAC-Nanos3-mRFP(Nos3-3′UTR) (blue in D) and BAC-Nanos3-mRFP(BghpA) (green in D) transgenic embryos at E7.5, E9.5 and E13.5. Data were normalized by G3PDH in each sample. The relative mRNA levels in the E9.5A sample (an anterior part of E9.5 embryo) were assigned the reference value of 1.0. A, anterior part of the embryo; P, posterior part of the embryo; M–K or K, the male kidney; M–G or G, the male gonad; F–G, the female gonad; N, the posterior part of a Nanos3 knockout embryo at E9.5. Error bars represent the s.d.

Mentions: We next examined the endogenous Nanos3 mRNA levels in somatic tissues between E9.5 and E13.5 by quantitative RT-PCR (qRT-PCR) and found transcripts even in somatic tissues (Fig. 4A–C), consistent with the above data. The anterior half of the embryo at E9.5 (9.5A) and the kidney at E13.5 (13.5K) do not contain any germ cells, but Nanos3 mRNA was detected (Fig. 4B–C). The level of Nanos3 mRNA in somatic tissue from E9.5 to E13.5 was maintained at very low levels compared with the gonads which containing many germ cells (Fig. 4C). Interestingly, Nanos3 expression was not detected in the anterior half of the embryo at E7.5 (E7.5A), suggesting the transcription of this gene is restricted to the PGCs when they are formed and may be slightly increased in the somatic cells at the later stage. It is consistent with previous reports, which include single–cell PCR analyses demonstrating Nanos3 expression exclusively in the PGCs [33] and our lineage study using Nanos3-cre [30].


The Nanos3-3'UTR is required for germ cell specific NANOS3 expression in mouse embryos.

Suzuki H, Saba R, Sada A, Saga Y - PLoS ONE (2010)

The accumulation of somatic Nanos3 mRNA is suppressed by the Nanos3-3′UTR.The levels of Nanos3 (A–C) or mRFP (D) mRNA were compared by quantitative RT-PCR using RNA samples derived from wild-type embryos (A–C) and BAC-Nanos3-mRFP(Nos3-3′UTR) (blue in D) and BAC-Nanos3-mRFP(BghpA) (green in D) transgenic embryos at E7.5, E9.5 and E13.5. Data were normalized by G3PDH in each sample. The relative mRNA levels in the E9.5A sample (an anterior part of E9.5 embryo) were assigned the reference value of 1.0. A, anterior part of the embryo; P, posterior part of the embryo; M–K or K, the male kidney; M–G or G, the male gonad; F–G, the female gonad; N, the posterior part of a Nanos3 knockout embryo at E9.5. Error bars represent the s.d.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2823788&req=5

pone-0009300-g004: The accumulation of somatic Nanos3 mRNA is suppressed by the Nanos3-3′UTR.The levels of Nanos3 (A–C) or mRFP (D) mRNA were compared by quantitative RT-PCR using RNA samples derived from wild-type embryos (A–C) and BAC-Nanos3-mRFP(Nos3-3′UTR) (blue in D) and BAC-Nanos3-mRFP(BghpA) (green in D) transgenic embryos at E7.5, E9.5 and E13.5. Data were normalized by G3PDH in each sample. The relative mRNA levels in the E9.5A sample (an anterior part of E9.5 embryo) were assigned the reference value of 1.0. A, anterior part of the embryo; P, posterior part of the embryo; M–K or K, the male kidney; M–G or G, the male gonad; F–G, the female gonad; N, the posterior part of a Nanos3 knockout embryo at E9.5. Error bars represent the s.d.
Mentions: We next examined the endogenous Nanos3 mRNA levels in somatic tissues between E9.5 and E13.5 by quantitative RT-PCR (qRT-PCR) and found transcripts even in somatic tissues (Fig. 4A–C), consistent with the above data. The anterior half of the embryo at E9.5 (9.5A) and the kidney at E13.5 (13.5K) do not contain any germ cells, but Nanos3 mRNA was detected (Fig. 4B–C). The level of Nanos3 mRNA in somatic tissue from E9.5 to E13.5 was maintained at very low levels compared with the gonads which containing many germ cells (Fig. 4C). Interestingly, Nanos3 expression was not detected in the anterior half of the embryo at E7.5 (E7.5A), suggesting the transcription of this gene is restricted to the PGCs when they are formed and may be slightly increased in the somatic cells at the later stage. It is consistent with previous reports, which include single–cell PCR analyses demonstrating Nanos3 expression exclusively in the PGCs [33] and our lineage study using Nanos3-cre [30].

Bottom Line: This is fundamental to the continuation of a species.Although Nanos3 is transcribed in both cell lineages, it is efficiently translated only in the germ lineage.Surprisingly, even when under the control of the CAG promoter which induces strong ubiquitous transcription in both germ cells and somatic cells, the addition of the Nanos3-3'UTR sequence to the coding region of exogenous gene was effective in restricting protein expression in germ cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, Graduate School of Science, University of Tokyo, Tokyo, Japan.

ABSTRACT

Background: The regulation of gene expression via a 3' untranslated region (UTR) plays essential roles in the discrimination of the germ cell lineage from somatic cells during embryogenesis. This is fundamental to the continuation of a species. Mouse NANOS3 is an essential protein required for the germ cell maintenance and is specifically expressed in these cells. However, the regulatory mechanisms that restrict the expression of this gene in the germ cells is largely unknown at present.

Methodology/principal findings: In our current study, we show that differences in the stability of Nanos3 mRNA between germ cells and somatic cells is brought about in a 3'UTR-dependent manner in mouse embryos. Although Nanos3 is transcribed in both cell lineages, it is efficiently translated only in the germ lineage. We also find that the translational suppression of NANOS3 in somatic cells is caused by a 3'UTR-mediated mRNA destabilizing mechanism. Surprisingly, even when under the control of the CAG promoter which induces strong ubiquitous transcription in both germ cells and somatic cells, the addition of the Nanos3-3'UTR sequence to the coding region of exogenous gene was effective in restricting protein expression in germ cells.

Conclusions/significance: Our current study thus suggests that Nanos3-3'UTR has an essential role in translational control in the mouse embryo.

Show MeSH
Related in: MedlinePlus