Olfactory receptor mRNAs act as selfish non-coding RNAs that enforce transcriptional singularity
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ABSTRACT:
PROVIDER: PRJNA960870 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR24128684.fastq.gz | Fastqsanger.gz | |||
| SRR24128685_1.fastq.gz | Fastqsanger.gz | |||
| SRR24128685_2.fastq.gz | Fastqsanger.gz | |||
| SRR24128686_1.fastq.gz | Fastqsanger.gz | |||
| SRR24128686_2.fastq.gz | Fastqsanger.gz |
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Project description:Olfactory receptor mRNAs act as selfish non-coding RNAs that enforce transcriptional singularity (OR Switching Experiment)
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Project description:To investigate roles for olfactory receptor coding sequence mRNA in trans olfactory receptor gene silencing, and OR-gene enhancer recruitment. Olfactory receptor (OR) choice provides an extreme example of allelic competition for transcriptional dominance, as every olfactory neuron stably transcribes one out of more than ~2,000 OR alleles1,2. OR gene choice is mediated by a multi-chromosomal enhancer hub that activates transcription at a single OR3,4, followed by ORtranslation dependent feedback that stabilizes this choice5,6. However, with more than one OR enhancer hub in each nucleus7, the mechanism by which only one OR allele is chosen for transcription remains enigmatic. Here, using single cell genomics, we show that only one of the OR enhancer hubs formed in each neuron retains euchromatic features and sustains transcriptional competence. Further, we provide evidence that OR transcription recruits enhancers and reinforces enhancer hub activity locally, while OR RNA inhibits transcription in competing hubs over distance, promoting OR protein-independent transition to singularity. While OR RNA synthesis is sufficient to break the symmetry between equipotent enhancer hubs, OR translation stabilizes transcription at the prevailing hub, revealing dual non-coding and coding mechanisms implemented by OR RNAs for transcriptional prevalence. We propose that coding mRNAs can function as “selfish” non-coding RNAs that influence nuclear architecture, enhance their own transcription, and inhibit their competitors, with generalizable implications in probabilistic cell fate decisions.
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Project description:To investigate roles for olfactory receptor coding sequence mRNA in trans olfactory receptor gene silencing, and OR-gene enhancer recruitment. Olfactory receptor (OR) choice provides an extreme example of allelic competition for transcriptional dominance, as every olfactory neuron stably transcribes one out of more than ~2,000 OR alleles1,2. OR gene choice is mediated by a multi-chromosomal enhancer hub that activates transcription at a single OR3,4, followed by ORtranslation dependent feedback that stabilizes this choice5,6. However, with more than one OR enhancer hub in each nucleus7, the mechanism by which only one OR allele is chosen for transcription remains enigmatic. Here, using single cell genomics, we show that only one of the OR enhancer hubs formed in each neuron retains euchromatic features and sustains transcriptional competence. Further, we provide evidence that OR transcription recruits enhancers and reinforces enhancer hub activity locally, while OR RNA inhibits transcription in competing hubs over distance, promoting OR protein-independent transition to singularity. While OR RNA synthesis is sufficient to break the symmetry between equipotent enhancer hubs, OR translation stabilizes transcription at the prevailing hub, revealing dual non-coding and coding mechanisms implemented by OR RNAs for transcriptional prevalence. We propose that coding mRNAs can function as “selfish” non-coding RNAs that influence nuclear architecture, enhance their own transcription, and inhibit their competitors, with generalizable implications in probabilistic cell fate decisions.
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Project description:Olfactory receptor (Olfr) genes comprise the largest gene family in mice. Despite their importance, most Olfr mRNAs are uncharacterized. Using RNA-seq analysis, we annotated the repertoire of mouse Olfr mRNAs expressed in the olfactory epithelium and found that they have several atypical features suggesting how post-transcriptional regulation impacts their expression. First, many Olfr mRNAs are intronless and those with introns typically have a single intron at the 5’ end. Second, Olfr mRNAs, as a group, have dramatically higher average AU content and lower predicted secondary structure than do control mRNAs. Third, Olfr mRNAs have a higher density of AU-rich elements (AREs) than control mRNAs. Fourth, Olfr mRNAs have much shorter 3’ UTR regions and fewer predicted binding sites for neurally expressed miRNAs than control mRNAs. Fifth, Olfr mRNAs have a significantly greater number of upstream open reading frames (uORFs) in their 5’ UTR than control mRNAs. All of these novel properties correlated with higher Olfr expression. Finally, we identified striking differences in the mRNAs from the two classes of Olfr genes, a finding consistent with their independent evolutionary origin. Our study suggests that the Olfr gene family has encountered unusual selective forces that have driven these unique post-transcriptional regulatory features. Examine transcriptome of Olfr genes in mouse olfactory epithelium
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Project description:Olfactory receptor (Olfr) genes comprise the largest gene family in mice. Despite their importance, most Olfr mRNAs are uncharacterized. Using RNA-seq analysis, we annotated the repertoire of mouse Olfr mRNAs expressed in the olfactory epithelium and found that they have several atypical features suggesting how post-transcriptional regulation impacts their expression. First, many Olfr mRNAs are intronless and those with introns typically have a single intron at the 5’ end. Second, Olfr mRNAs, as a group, have dramatically higher average AU content and lower predicted secondary structure than do control mRNAs. Third, Olfr mRNAs have a higher density of AU-rich elements (AREs) than control mRNAs. Fourth, Olfr mRNAs have much shorter 3’ UTR regions and fewer predicted binding sites for neurally expressed miRNAs than control mRNAs. Fifth, Olfr mRNAs have a significantly greater number of upstream open reading frames (uORFs) in their 5’ UTR than control mRNAs. All of these novel properties correlated with higher Olfr expression. Finally, we identified striking differences in the mRNAs from the two classes of Olfr genes, a finding consistent with their independent evolutionary origin. Our study suggests that the Olfr gene family has encountered unusual selective forces that have driven these unique post-transcriptional regulatory features.
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Project description:Olfactory ensheathing cells are one of the few central nervous system regenerative cells discovered so far. It is characterized by its lifelong nerve regeneration function, and it can also release a variety of neurotrophic factors and neural adhesion molecules. It is considered to be the glial cell with the strongest myelination ability. Olfactory ensheathing cells and Schwann cells have phenotypes in common, they can promote axon regeneration(R. Doucette, 1995). Olfactory ensheathing cells have the characteristics of Schwann cells and astrocytes, but the overall performance tends to be the former, which has two unique characteristics. First, it exists not only in the peripheral nerves (Schwann cells), but also in the central nervous system (astroglia); second, the olfactory mucosa has the ability to regenerate life-long, including human olfactory ensheathing cells(J. C. Bartolomei and C. A. Greer, 2000). Regeneration is a process in which olfactory ensheathing cells participate in efficient regulation, although the specific mechanism is not yet clear. Olfactory ensheathing cells are different from astrocytes and Schwann cells, but at the same time have the characteristics of these two cells(S. C. Barnett, 2004), like Schwann cells help axon growth, but more than Schwann cells It can make axons grow long distances, that is, it has stronger migration(A. Ramon-Cueto et al., 1998); there are also astrocytes that have a nutritional effect on the survival of neurons and the growth of axons, but olfactory ensheathing cells can also wrap neurons forms myelin sheath to support the growth of nerve processes(R. Devon and R. Doucette, 1992; J. Gu et al., 2019). There are two characteristics that make olfactory ensheathing cells the best choice for the treatment of neurological diseases(S. C. Chiu et al., 2009; J. Kim et al., 2018; M. Abdel-Rahman et al., 2018). Olfactory ensheathing cells are gradually used to treat spinal cord injuries and have shown amazing effects(J. C. Bartolomei and C. A. Greer, 2000; K. J. Liu et al., 2010; R. Yao et al., 2018). Olfactory ensheathing cells that have been used in research are usually derived from the olfactory bulb(E. H. Franssen et al., 2007), but it is easier to obtain olfactory ensheathing cells from the olfactory mucosa in clinical practice(M. Ryszard et al., 2006), so the difference between the olfactory ensheathing cells from the olfactory bulb and the olfactory mucosa There are more and more studies(B. M. U. et al., 2007), and previous studies have shown that they not only have many similar functions, but also have many differences(M. W. Richter et al., 2005; L. Wang et al., 2014; K. E. Smith et al., 2020). Because olfactory ensheathing cells derived from the olfactory bulb are not easy to obtain, olfactory ensheathing cells derived from the olfactory mucosa have become the focus of attention. Although we know that olfactory ensheathing cells from two sources have nerve repair functions, it is not clear why the two different sources of olfactory ensheathing cells have different therapeutic effects. Nicolas G. once studied that the genetic difference between the two cells and found that there are many genes related to wound repair and nerve regeneration(G. Nicolas et al., 2010). We have reason to guess that olfactory ensheathing cells from these two sources will also have a large difference in protein level. Our research group wants to use the current mature transcriptome and proteomic sequencing technologies to explore the difference between olfactory ensheathing cells from the olfactory bulb and olfactory mucosa, and explain why the two sources of olfactory ensheathing cells shows different therapeutic effects, hope to provide a new theoretical basis for future clinical treatment.
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