Dataset Information

Comparative analyses of vertebrate CPEB-family of proteins define two sub-families with distinct, but coordinated, functions in post-transcriptional regulation of gene expression

ABSTRACT: Background: The vertebrate CPEB-family of RNA-binding proteins promote translational repression or activation of their target mRNAs, which harbor cytoplasmic polyadenylation elements (CPEs) in their 3’ UTRs, through cytoplasmic changes in their poly(A) tail lengths. However, their regulation(s) and mechanism(s) of action have not been systematically addressed as a family. Results: Here we perform a comparative analysis of the four vertebrate CPEBs in their regulation by phosphorylation, supramolecular assemblies’ composition and properties and in their mRNA targets. Conclusions: We show that although all four CPEBs are able to recruit CCR4-NOT deadenylation complex when not phosphorylated, their mechanisms of action determine two subfamilies with distinct, but coordinated, regulation by phosphorylation and target specificity. Thus, CPEB1 forms ribonucleoprotein complexes that are remodeled upon a single phosphorylation event, by AurkA, and are associated with mRNAs containing canonical CPEs. On the other hand, CPEB2-4 are regulated by multiple proline-directed phosphorylations that control their liquid-liquid phase-separation. CPEB2-4 mRNA-targets include CPEB1-bound transcripts, with canonical CPEs, but also a specific subset of mRNAs with non-canonical CPEs. In turn, CPEB2-4 coacervates display compositional, morphological and dynamic differences. Altogether, these results show how, globally, the CPEB family of proteins is able to integrate cellular cues to generate a finetuned adaptive response in gene expression regulation.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Xenopus Laevis (african Clawed Frog)

TISSUE(S): Oocyte

SUBMITTER: Marina Gay

LAB HEAD: Raul Mendez

PROVIDER: PXD030419 | Pride | 2022-10-05

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	2591_MC_10_BirA_RBD_ZZ_F2_SCX.raw	Raw
	2591_MC_11_RBD_ZZ_BirA_F2_SCX.raw	Raw
	2591_MC_12_BirA_MS2_F2_SCX.raw	Raw
	2591_MC_13_BirA_F3_SCX.raw	Raw
	2591_MC_14_BirA_xCPEB1_F3_SCX.raw	Raw

Items per page:

1 - 5 of 110

Publications

Comparative analyses of vertebrate CPEB proteins define two subfamilies with coordinated yet distinct functions in post-transcriptional gene regulation.

Duran-Arqué Berta B Cañete Manuel M Castellazzi Chiara Lara CL Bartomeu Anna A Ferrer-Caelles Anna A Reina Oscar O Caballé Adrià A Gay Marina M Arauz-Garofalo Gianluca G Belloc Eulalia E Mendez Raúl R Mendez Raúl R

Genome biology 20220912 1

<h4>Background</h4>Vertebrate CPEB proteins bind mRNAs at cytoplasmic polyadenylation elements (CPEs) in their 3' UTRs, leading to cytoplasmic changes in their poly(A) tail lengths; this can promote translational repression or activation of the mRNA. However, neither the regulation nor the mechanisms of action of the CPEB family per se have been systematically addressed to date.<h4>Results</h4>Based on a comparative analysis of the four vertebrate CPEBs, we determine their differential regulatio ...[more]

PMID: 36096799

Similar Datasets

Project description:An important facet of the oocyte to embryo transition in mammals is the rapid elimination of a subset of the maternal products that got accumulated during oogenesis. RNA studies support a view that the transition occurs quite rapidly. Whether this applies also for proteins remains to be determined beyond the very few cases known to date. We report that COPS3, the 3rd subunit of the COP9 signalosome complex, forms a large protein deposit in mouse oocytes (94th percentile of the riBAQ distribution). The one and only knock-out study had previously shown that Cops3 null (-/-) mouse embryos obtained from heterozygous intercrosses arrested after 5.5 dpc and were resorbed by 8.5 dpc mainly due to increased cell death in the epiblast (PMID: 12972600). However, more recent studies from our group ascribed Cops3 gene with a developmental role already at the 2-cell stage. Specifically, the sister blastomeres differ from each other in Cops3 mRNA levels and distribution, preceding the discordance of epiblast formation in derivative twin blastocysts, and implicating Cops3 in the molecular underpinnings of totipotency. This discrepancy between original knock-out result and recent observations can be resolved if we posit that the first requirement for Cops3 gene function was bridged by maternal protein that outlived the locus excision, consistent with the observation that Cops3 -/- blastocysts had the same immunostaining intensity as the +/- or +/+ counterparts in the original knock-out study. Thus, these contradicting observations support a hypothesis that 5.5 dpc may not have been the first time when the gene function was needed in development, but the second time. To test this hypothesis, pronuclear-stage mouse oocytes were subjected to an immunological method that directly targets the protein of interest by way of proteasomal degradation of the COPS3-antibody-TRIM21 complex. Briefly, pronuclear-stage mouse oocytes were microinjected with a mixture of the COPS3-specific purified monoclonal IgG antibody, mCherry-Trim21 mRNA and Oregon Green dextran beads (inert tracer). Injected oocytes were then allowed to develop and collected for lysis 24 hours later, when they reached the 2-cell stage but were not able to progress further. In addition to the COPS3, we targeted two additional proteins, OCT4 and TEAD4. The following seven experimental groups were examined by liquid chromatography-mass spectrometry (LC-MS/MS) using the pipeline described previously by Israel et al.: 1) non-manipulated 2-cell embryos; 2) 2-cell embryos from oocytes injected with Oregon Green dextran beads; 3) 2-cell embryos from oocytes injected with mCherry-Trim21 mRNA and Oregon Green; 4) 2-cell embryos from oocytes injected with mCherry-Trim21 mRNA, Oregon Green and anti-COPS3 (Abcam 79698); 5) 2-cell embryos from oocytes injected with anti-COPS3 (Abcam 79698) and Oregon Green; 6) 2-cell embryos from oocytes injected with mCherry-Trim21 mRNA, Oregon Green and anti-TEAD4 (Abcam 58310); 7) 2-cell embryos from oocytes injected with mCherry-Trim21 mRNA, Oregon Green and anti-OCT4 (Abcam 181557).