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Status |
Public on Sep 01, 2020 |
Title |
Ancestral function of Inhibitors-of-kappaB regulates Caenorhabditis elegans development (RNA-Seq) |
Organism |
Caenorhabditis elegans |
Experiment type |
Expression profiling by high throughput sequencing
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Summary |
NF-κB-mediated signaling is maintained silent by the action of IκB proteins, whose canonical role is to sequester NF-κB in the cytoplasm. An alternative chromatin role for IκB members have been shown to affect stemness and cell differentiation but the involvement of NF-κB in this function has not been excluded. NFKI-1 and IKB-1 are IκB homologs in Caenorhabditis elegans, which lacks NF-κB nuclear effectors. nfki-1 and ikb-1 mutants present developmental defects that phenocopy mutations in Polycomb genes and demethylases as utx-1. suggesting a role for C. elegans IKB proteins in chromatin regulation, which is supported by various lines of evidence:(i) we detected NFKI-1 in the nucleus; (ii) NFKI-1 and IKB-1 bind to histones and Polycomb proteins, (iii) NFKI-1 and IKB-1 bind to chromatin in vivo, and (iv) mutations in nfki-1 and ikb-1 alter chromatin marks . Thus, ancestral IκB inhibitors may exert nuclear functions regulating of gene expression and development.
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Overall design |
To study the transcriptional alterations in nfki-1 and ikb-1 mutants during animal development, we performed RNA-seq at two different timepoints: L1 (5 hours fed and 3 hours starved at 25ºC after synchronization, 1 replicate per strain) and L4 (36 hours at 25ºC after synchronization, 2 replicates per strain) developmental stages. We used different mutant strains for nfki-1 and ikb-1 to avoid mutant-specific alterations. cer1 allele is a CRISPR-generated 368 bp deletion near nfki-1 exon 2. cer2 is a 434 bp deletion and 50 bp insertion at exon 2. Both modifications generate premature stop codons. cer9 allele is a 462 bp deletion that includes the ATG start codon of ikb-1. nr2027 is a 1911 bp deletion of ikb-1 that includes the ankyrin-repeat domains encoding sequence (Pujol et al., 2001). N2(Bristol) was used for all analyses as wildtype (WT) control. 50 bp single-end sequencing was done on an Illumina HiSeq 2000.
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Contributor(s) |
Brena D, Porta-de-la-Riva M, Bertran J, Guillén Y, Espinosa L, Cerón J |
Citation(s) |
32999373 |
Submission date |
Mar 09, 2020 |
Last update date |
Oct 06, 2020 |
Contact name |
Yolanda Guillén |
E-mail(s) |
yguillen87@gmail.com
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Organization name |
IMIM
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Street address |
Dr. Aiguader 88 Planta 2 IMIM
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City |
Barcelona |
ZIP/Postal code |
08003 |
Country |
Spain |
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Platforms (1) |
GPL13657 |
Illumina HiSeq 2000 (Caenorhabditis elegans) |
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Samples (14)
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This SubSeries is part of SuperSeries: |
GSE146655 |
Ancestral function of Inhibitors-of-kappaB regulates Caenorhabditis elegans development |
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Relations |
BioProject |
PRJNA611580 |
SRA |
SRP252085 |