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Methylation of the guanosine cap structure at the 5' end of mRNA is essential for efficient translation of all eukaryotic cellular mRNAs, gene expression and cell viability and promotes transcription, splicing, polyadenylation and nuclear export of mRNA. In the current study, we present the spatial expression pattern of the Xenopus laevis rnmt homologue. A high percentage of protein sequence similarity, especially within the methyltransferase domain, as well as an increased expression in the cells of the transcriptionally active stages, suggests a conserved RNA cap methylation function. Spatial expression analysis identified expression domains in the brain, the retina, the lens, the otic vesicles and the branchial arches.
Fig. 2.Temporal expression and comparison of rnmt paralogs. (A) The nucleotide sequence of the fragment used to compare spatial expression of rnmt-a and -b is shown. Sequences used for primer pairs are in red. Intron spanning primers were used to exclude any contamination from genomic DNA. Primer pairs, specific to each of both loci were used to compare the temporal expression of the paralogs.
Figure 2. (continued) (B) Semi-quantitative rt-PCR revealed zygotic expression of rnmt-a, whereas rnmt-b shows maternal presence. Housekeeping gene ODC1 was used as a control for equal RNA input. (C) Genomic profile of both the loci ‘a’ and ‘b’. Grey boxes indicate exons and lines joining them the intronic regions of the gene. Forward arrow repre-
sent the start codon while star the stop codon. Arrowheads over exons represent the PCR primer pairs used in this study. Note the missing genomic sequence is shown in dotted lines between exon 1 and exon 3 and including the whole exon.
rnmt (RNA (guanine-7-) methyltransferase) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 10, animal view (A) and vegetal view(B).
rnmt (RNA (guanine-7-) methyltransferase) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 12, animal view (C) and vegetal view (D).
rnmt (RNA (guanine-7-) methyltransferase) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 15, anterio-dorsal view, anterior down.
rnmt (RNA (guanine-7-) methyltransferase) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 24,(I): lateral view, anteriorleft, dorsal up, and( J) anterior view, dorsal up.
rnmt (RNA (guanine-7-) methyltransferase) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 27 (L): lateral view, anteriorleft, dorsal up, and (M) anterior view, dorsal up.
