FIG. 4. Upstream AUGs down-regulate translation of the long SHOX 5′-UTR in vitro and in vivo. A, schematic of the in vitro-transcribed mRNAs used to examine the impact of uAUGs on translation efficiency. The 5′-UTRs of these mRNAs are identical in size and exhibit comparable structural features as determined by the free energy values for their predicted folding. B, translation efficiency of in vitro mutagenized RNAs after transfection into U2Os cells. Translation attenuating effects of the type 1 transcripts can be attributed to uAUGs present in their 5′-UTR because mutations of uAUG3–7 or all seven uAUGs restore translation activities comparable with type 2 transcripts. C, schematic and gelelectrophoretic analysis of UTR-noggin fusion RNAs used for Xenopus embryo injection. Type 1 UTR (1), type 2 UTR (2), and mutagenized type 1 UTR (3) are fused to a noggin cDNA. D, these mRNAs yield three distinct dorsalized phenotypes. The weakest dorsalizing effect was observed with type 1 transcripts (1, wild type), whereas injection of both, type 2 (2), and mutagenized type 1 transcripts (3) resulted in embryos with more severe trunk and tail defects. Injection of increasing amounts (50, 100, and 500 pg) of different transcripts (1, 2, and 3) result in stronger dorsalization of the embryos. The phenotypes are characterized as wild type (blue), moderately (red), and strongly dorsalized (yellow). The effects of all three mRNAs exhibit a typical dose respose.
Image published in: Blaschke RJ et al. (2003)
Copyright © 2003. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License.
| Experiment + Assay | Source | Phenotypes and Disease | ||
|---|---|---|---|---|
| Xla Wt + Hsa.SHOX-nog + NF33/34-35/36 (whole-mount microscopy) | Fig.4.D |
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| Xla Wt + Hsa.SHOXdel-nog + NF33/34-35/36 (whole-mount microscopy) | Fig.4.D |
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| Xla Wt + Hsa.SHOXdel-nog + NF33/34-35/36 (whole-mount microscopy) | Fig.4.D |
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| Xla Wt + Hsa.SHOXtm-nog + NF33/34-35/36 (whole-mount microscopy) | Fig.4.D |
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| Xla Wt + Hsa.SHOXtm-nog + NF33/34-35/36 (whole-mount microscopy) | Fig.4.D |
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