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. 2003 Feb;9(2):180-6.
doi: 10.1261/rna.2141503.

Numerous microRNPs in neuronal cells containing novel microRNAs

Josée Dostie  1 , Zissimos Mourelatos, Michael Yang, Anup Sharma, Gideon Dreyfuss
Affiliations

Numerous microRNPs in neuronal cells containing novel microRNAs

Josée Dostie et al. RNA. 2003 Feb.

Erratum in

  • RNA. 2003 May;9(5):631-2

Abstract

Spinal muscular atrophy (SMA) is a common neurodegenerative disease that is caused by deletions or loss-of-function mutations in the Survival of Motor Neuron (SMN) protein. SMN is part of a large complex that functions in the assembly/restructuring of ribonucleoprotein (RNP) complexes. We recently showed in HeLa cells that two components of the SMN complex, Gemin3 and Gemin4, together with the argonaute protein eIF2C2, also associate with microRNAs (miRNAs) as part of a novel class of RNPs termed miRNPs. Here we report on miRNPs isolated from neuronal cell lines of mouse and human, and describe 53 novel miRNAs. Several of these miRNAs are conserved in divergent organisms, including rat, zebrafish, pufferfish, and the nematode Caenorhabditis elegans. The chromosomal locations of most of the novel miRNAs were identified and indicate some phylogenetic conservation of the likely precursor structures. Interestingly the gene locus of one miRNA, miR-175, is a candidate region for two neurologic diseases: early-onset parkinsonism (Waisman syndrome) and X-linked mental retardation (MRX3). Also, several miRNAs identified as part of miRNPs in these cells appear to constitute two distinct subfamilies. These subfamilies comprise multiple copies of miRNAs on different chromosomes, suggesting an important function in the regulation of gene expression.

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Figures

FIGURE 1.
FIGURE 1.
miRNPs from mouse and human neuronal cells. Total extracts from MN-1 (mouse) and Weri (human) cells were incubated with either nonimmune mouse serum (NMS) as a control or the anti-Gemin3 antibody (11G9). Immunoprecipitates were treated with proteinase K, and associated RNAs were purified by phenol-chloroform extraction and precipitated. Isolated RNAs were 3′-end-labeled with [5′-32P]-pCp and resolved by electrophoresis on 15% denaturing polyacrylamide gels. Left: molecular weight marker.
FIGURE 2.
FIGURE 2.
Predicted secondary structures of the novel human miRNA putative precursors. Human genomic sequences upstream and downstream of the novel miRNAs were folded with the computer program mfold. Predicted secondary structures are shown 5′ to 3′. Red areas represent the mature miRNA. Chromosome number represents the localization of the miRNA in the human genome.
FIGURE 3.
FIGURE 3.
Novel miRNA clusters and families of miRNA paralogs associated with miRNPs in neuronal cells. (A) Novel miRNA clusters. The putative precursors of identified miRNAs are presented as solid boxes, and the mature miRNAs identified in the study are shown in red. Computer-predicted putative miRNA precursors are presented as dashed boxes, and the predicted mature miRNAs are shown in light blue. The chromosomal localization in the human genome is indicated on the right. The size of the region containing the miRNAs is indicated at the top of each cluster. (B) miRNPs contain a conserved miRNA family in neuronal cells. miRNA family members associated with miRNPs in neuronal cells are aligned. The names of miRNAs are indicated on the left. miRNA length is shown at the end of each miRNA. Conserved residues are in bold letters and gray shaded areas. Consensus is shown at the bottom: Nucleotides shown in red were invariable. (C) miRNPs contain a class of miRNAs with homologous 5′ ends in neuronal cells. miRNA class members with homologous 5′ ends are aligned. The names of miRNAs are indicated on the left. miRNA length is shown at the end of each miRNA. Conserved residues are in bold letters and gray shaded areas. Consensus is shown at the bottom: Nucleotides shown in red were invariable.
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