Fig. 1: mamRNA is produced from a U14-containing precursor and undergoes splicing. | Nature Communications

Fig. 1: mamRNA is produced from a U14-containing precursor and undergoes splicing.

From: A bifunctional snoRNA with separable activities in guiding rRNA 2’-O-methylation and scaffolding gametogenesis effectors

Fig. 1

a Total and poly(A) + RNA-seq profiles of the mamRNA and U14-encoding genomic locus (n = 2). The numbers above the scheme indicate the nucleotide length from mamRNA TSS (black arrow). The numbers in square brackets indicate the range of reads. Below are shown the sequences at the borders of the intron with the 5’ splice site (5’SS), branchpoint (BP) and 3’ splice site (3’SS) highlighted in bold. Nucleotides mutated in dedicated strains are shown below in red. b Semi-quantitative RT-PCR assay of the mamRNA-U14 locus. Left, scheme showing the different strains used, with point mutations labeled with red stars. The primers used are denoted by green arrows. Right, cDNA analysis on agarose gels. act1+ was used as a loading control and no RT reactions (-RT) were loaded in parallel. bp = base pairs. c Northern blots showing mamRNA and U14 levels from total RNA samples, in the indicated genetic backgrounds. The position of the different probes used (A, B, C and D) are depicted on top. Ribosomal RNAs served as loading control (lower panels). b = bases. d and e Representative images of mamRNA localization in mitotic cells of the indicated genetic backgrounds, expressing either GFP-tagged Mmi1 or Nop56. smFISH probes used were specific of mamRNA 5’ exon and DNA was stained with DAPI. Images are shown as Z-projections. Scale bar, 5 µm. In d white arrows point to Mmi1 dot-mamRNA colocalization. Source data are provided as a Source Data file.

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