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Decreased Expression of CoREST1 and CoREST2 Together with LSD1 and HDAC1/2 during Neuronal Differentiation.

Sáez JE, Gómez AV, Barrios ÁP, Parada GE, Galdames L, González M, Andrés ME - PLoS ONE (2015)

Bottom Line: In both models, a concomitant and gradual decrease of LSD1, HDAC1, HDAC2, CoREST1, and CoREST2, but not CoREST3 was observed.In conclusion, neuronal differentiation is accompanied by decreased expression of all core components of LCH complexes, but not CoREST3.The combination of the differential transcriptional repressor capacity of LCH complexes and variable protein levels of its different components should result in a finely tuned gene expression during neuronal differentiation and in the adult brain.

View Article: PubMed Central - PubMed

Affiliation: Department of Cellular and Molecular Biology, Faculty of Biological Sciences, Pontificia Universidad Católica de Chile, Santiago, Chile.

ABSTRACT
CoREST (CoREST1, rcor1) transcriptional corepressor together with the histone demethylase LSD1 (KDM1A) and the histone deacetylases HDAC1/2 form LSD1-CoREST-HDAC (LCH) transcriptional complexes to regulate gene expression. CoREST1 belong to a family that also comprises CoREST2 (rcor2) and CoREST3 (rcor3). CoREST1 represses the expression of neuronal genes during neuronal differentiation. However, the role of paralogs CoREST2 and CoREST3 in this process is just starting to emerge. Here, we report the expression of all CoRESTs and partners LSD1 and HDAC1/2 in two models of neuronal differentiation: Nerve-Growth-Factor (NGF)-induced neuronal phenotype of PC12 cells, and in vitro maturation of embryonic rat cortical neurons. In both models, a concomitant and gradual decrease of LSD1, HDAC1, HDAC2, CoREST1, and CoREST2, but not CoREST3 was observed. As required by the study, full-length rat rcor1 gene was identified using in silico analysis of available rat genome. The work was also complemented by the analysis of rat RNA-seq databases. The analysis showed that all CoRESTs, including the identified four splicing variants of rat CoREST3, display a wide expression in adult tissues. Moreover, the analysis of RNA-seq databases showed that CoREST2 displays a higher expression than CoREST1 and CoREST3 in the mature brain. Immunofluorescent assays and immunoblots of adult rat brain showed that all CoRESTs are present in both glia and neurons. Regarding functional partnership, CoREST2 and CoREST3 interact with all LSD1 splicing variants. In conclusion, neuronal differentiation is accompanied by decreased expression of all core components of LCH complexes, but not CoREST3. The combination of the differential transcriptional repressor capacity of LCH complexes and variable protein levels of its different components should result in a finely tuned gene expression during neuronal differentiation and in the adult brain.

No MeSH data available.


Annotation of rat rcor1-3 transcripts.UCSC Genome Browser images show transcript annotation data at rcor1 (A), rcor2 (B) and rcor3 (C) loci. Gene model tracks of RefSeq and Ensembl are shown in blue and red, respectively. Assembled transcripts by Cufflinks are shown in black. Predicted ELM2 and SANT1/SANT2 domains are highlighted in yellow and red, respectively. Together, the alignments of RefSeq transcripts annotation from human and mouse with the PhyloP genomic conservation scores give the evolutionary information to support the existence of the new identified exons.
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pone.0131760.g001: Annotation of rat rcor1-3 transcripts.UCSC Genome Browser images show transcript annotation data at rcor1 (A), rcor2 (B) and rcor3 (C) loci. Gene model tracks of RefSeq and Ensembl are shown in blue and red, respectively. Assembled transcripts by Cufflinks are shown in black. Predicted ELM2 and SANT1/SANT2 domains are highlighted in yellow and red, respectively. Together, the alignments of RefSeq transcripts annotation from human and mouse with the PhyloP genomic conservation scores give the evolutionary information to support the existence of the new identified exons.

Mentions: Rat rcor1-3 transcripts expression was analyzed using public available RNA-seq data from eight different tissues [25]. Using Cufflinks [27] we assembled the RNA-seq read alignments at the rcor1-3 loci. The assembled transcripts showed a strong evolutionary correspondence with the rcor1-3 transcripts of mouse and human models (Fig 1A). At rcor1 locus, whereas RefSeq [30] track shows a single unspliced transcript, Ensembl [31] track shows a ten-exon transcript that is coincident with mouse and human rcor1 transcript model. Our transcript assembly of rcor1 allowed identifying two additional exons that were not annotated at Ensembl gene model. These additional exons codify for the ELM2 domain of rat CoREST1 (Fig 1A). At rcor2 locus, the assembled transcript is coincident with the rcor2 gene models of RefSeq and Ensembl (Fig 1B). Finally, at the rcor3 locus, we assembled three rcor3 isoforms that are not annotated in RefSeq or Ensembl. Interestingly, one of the novel isoforms (rcor3-b) contains three unannotated exons, and two of them codify for an additional SANT2 (SANT2-b) domain that is only included in this isoform. None of the new assembled rcor3 transcripts contains both SANT2 domains, suggesting that the inclusion of the pair of exons that codify for the SANT2 domains are mutually exclusive (Fig 1C). Duplication of SANT2 domains at rcor3 locus were not detected in human or mouse (S1 Fig).


Decreased Expression of CoREST1 and CoREST2 Together with LSD1 and HDAC1/2 during Neuronal Differentiation.

Sáez JE, Gómez AV, Barrios ÁP, Parada GE, Galdames L, González M, Andrés ME - PLoS ONE (2015)

Annotation of rat rcor1-3 transcripts.UCSC Genome Browser images show transcript annotation data at rcor1 (A), rcor2 (B) and rcor3 (C) loci. Gene model tracks of RefSeq and Ensembl are shown in blue and red, respectively. Assembled transcripts by Cufflinks are shown in black. Predicted ELM2 and SANT1/SANT2 domains are highlighted in yellow and red, respectively. Together, the alignments of RefSeq transcripts annotation from human and mouse with the PhyloP genomic conservation scores give the evolutionary information to support the existence of the new identified exons.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4482511&req=5

pone.0131760.g001: Annotation of rat rcor1-3 transcripts.UCSC Genome Browser images show transcript annotation data at rcor1 (A), rcor2 (B) and rcor3 (C) loci. Gene model tracks of RefSeq and Ensembl are shown in blue and red, respectively. Assembled transcripts by Cufflinks are shown in black. Predicted ELM2 and SANT1/SANT2 domains are highlighted in yellow and red, respectively. Together, the alignments of RefSeq transcripts annotation from human and mouse with the PhyloP genomic conservation scores give the evolutionary information to support the existence of the new identified exons.
Mentions: Rat rcor1-3 transcripts expression was analyzed using public available RNA-seq data from eight different tissues [25]. Using Cufflinks [27] we assembled the RNA-seq read alignments at the rcor1-3 loci. The assembled transcripts showed a strong evolutionary correspondence with the rcor1-3 transcripts of mouse and human models (Fig 1A). At rcor1 locus, whereas RefSeq [30] track shows a single unspliced transcript, Ensembl [31] track shows a ten-exon transcript that is coincident with mouse and human rcor1 transcript model. Our transcript assembly of rcor1 allowed identifying two additional exons that were not annotated at Ensembl gene model. These additional exons codify for the ELM2 domain of rat CoREST1 (Fig 1A). At rcor2 locus, the assembled transcript is coincident with the rcor2 gene models of RefSeq and Ensembl (Fig 1B). Finally, at the rcor3 locus, we assembled three rcor3 isoforms that are not annotated in RefSeq or Ensembl. Interestingly, one of the novel isoforms (rcor3-b) contains three unannotated exons, and two of them codify for an additional SANT2 (SANT2-b) domain that is only included in this isoform. None of the new assembled rcor3 transcripts contains both SANT2 domains, suggesting that the inclusion of the pair of exons that codify for the SANT2 domains are mutually exclusive (Fig 1C). Duplication of SANT2 domains at rcor3 locus were not detected in human or mouse (S1 Fig).

Bottom Line: In both models, a concomitant and gradual decrease of LSD1, HDAC1, HDAC2, CoREST1, and CoREST2, but not CoREST3 was observed.In conclusion, neuronal differentiation is accompanied by decreased expression of all core components of LCH complexes, but not CoREST3.The combination of the differential transcriptional repressor capacity of LCH complexes and variable protein levels of its different components should result in a finely tuned gene expression during neuronal differentiation and in the adult brain.

View Article: PubMed Central - PubMed

Affiliation: Department of Cellular and Molecular Biology, Faculty of Biological Sciences, Pontificia Universidad Católica de Chile, Santiago, Chile.

ABSTRACT
CoREST (CoREST1, rcor1) transcriptional corepressor together with the histone demethylase LSD1 (KDM1A) and the histone deacetylases HDAC1/2 form LSD1-CoREST-HDAC (LCH) transcriptional complexes to regulate gene expression. CoREST1 belong to a family that also comprises CoREST2 (rcor2) and CoREST3 (rcor3). CoREST1 represses the expression of neuronal genes during neuronal differentiation. However, the role of paralogs CoREST2 and CoREST3 in this process is just starting to emerge. Here, we report the expression of all CoRESTs and partners LSD1 and HDAC1/2 in two models of neuronal differentiation: Nerve-Growth-Factor (NGF)-induced neuronal phenotype of PC12 cells, and in vitro maturation of embryonic rat cortical neurons. In both models, a concomitant and gradual decrease of LSD1, HDAC1, HDAC2, CoREST1, and CoREST2, but not CoREST3 was observed. As required by the study, full-length rat rcor1 gene was identified using in silico analysis of available rat genome. The work was also complemented by the analysis of rat RNA-seq databases. The analysis showed that all CoRESTs, including the identified four splicing variants of rat CoREST3, display a wide expression in adult tissues. Moreover, the analysis of RNA-seq databases showed that CoREST2 displays a higher expression than CoREST1 and CoREST3 in the mature brain. Immunofluorescent assays and immunoblots of adult rat brain showed that all CoRESTs are present in both glia and neurons. Regarding functional partnership, CoREST2 and CoREST3 interact with all LSD1 splicing variants. In conclusion, neuronal differentiation is accompanied by decreased expression of all core components of LCH complexes, but not CoREST3. The combination of the differential transcriptional repressor capacity of LCH complexes and variable protein levels of its different components should result in a finely tuned gene expression during neuronal differentiation and in the adult brain.

No MeSH data available.