Castaldo, Daniela (2020) Identification and characterization of Lin28A molecular complexes regulating mRNA recognition and translation in Epiblast Stem Cells. [Tesi di dottorato]


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Item Type: Tesi di dottorato
Resource language: English
Title: Identification and characterization of Lin28A molecular complexes regulating mRNA recognition and translation in Epiblast Stem Cells
Date: 11 March 2020
Number of Pages: 88
Institution: Università degli Studi di Napoli Federico II
Department: Medicina Molecolare e Biotecnologie Mediche
Dottorato: Medicina molecolare e biotecnologie mediche
Ciclo di dottorato: 32
Coordinatore del Corso di dottorato:
Avvedimento, Vittorio
Date: 11 March 2020
Number of Pages: 88
Keywords: Protein Interactors
Settori scientifico-disciplinari del MIUR: Area 05 - Scienze biologiche > BIO/11 - Biologia molecolare
Date Deposited: 25 Mar 2020 10:21
Last Modified: 10 Nov 2021 09:53

Collection description

RNA binding protein Lin28A is involved in the first steps of mammalian development and in the reprogramming of adult cells into iPS cells. This protein is known to regulate let-7 miRNA family and several mRNAs. It is able to positively regulate some targets, such as Igf2 and Oct4 mRNA, or negatively, as in the case of Hmga2 mRNA. and let-7 miRNA. Since we observed Lin28A accumulation during the establishment of the embryonic stem cell (ESC) differentiated phenotype, we looked at new mRNA targets, coding for proteins involved in the differentiation of ESCs into epiblast stem cell and bearing Lin28 consensus sequence. We found Dnmt3A mRNA as new Lin28A target and its expression resulted to be positively regulated by Lin28A. Furthermore, we were wondering which are the molecular mechanisms underlying the post transcriptional regulation mediated by Lin28A. To do that, we developed an experimental setting to identify Lin28A partners, able to affect its function, based on size-exclusion chromatography, Lin28A co-immunoprecipitation and identification of co-immunoprecipitated proteins through mass spectrometry analysis. We preliminary evaluated the contribution of these putative interactors on the effect of Lin28A overexpression on Dnmt3A mRNA translation, through their silencing. We found proteins having a positive effect on Dnmt3A mRNA translation in a Lin28A-dependent manner and proteins having a negative effect on Dnmt3A mRNA translation that is counteracted by Lin28A. In conclusion, we assumed that some interactors could have an important role in Lin28A-mediated post transcriptional regulation of Dnmt3a expression. These preliminary evaluations provide the proof of concept that Lin28A is part of a molecular machinery, involving several proteins, that regulates the translation of Lin28A target mRNAs.


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