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TET2-mediated tRNA oxidation and its role in hematopoietic cancers

Applicant Gonskikh Yulia
Number 200056
Funding scheme Early Postdoc.Mobility
Research institution Departement für Chemie, Biochemie und Pharmazie Universität Bern
Institution of higher education Institution abroad - IACH
Main discipline Biochemistry
Start/End 01.01.2021 - 30.06.2022
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Keywords (1)

tRNA, epitranscriptomics, protein translation

Lay Summary (German)

Lead
Ten-Eleven Translocation-2 (TET2) ist eines der am häufigsten mutierten Gene bei mehreren Arten von Blutkrebs. Die Tumorsuppressor-Funktion des TET2 Enzyms wurde zuerst mit seiner Fähigkeit erklärt, Nukleotide der DNS zu oxidieren. Vor kurzem wurde gezeigt, dass TET2 auch RNS oxidieren kann. TET2 kann die Nukleotide der Transfer-RNS (tRNS) oxidieren and dadurch die Proteinbiosynthese verstärken.
Lay summary

Forschungsziel dieses Projektes ist, den molekularen Mechanismus zu bestimmen, der der verstärkten Proteinbiosynthese durch TET2-modifizierte tRNS zugrunde liegt, und seine Beteiligung an der Entwicklung von hämatopoetischen Krebsarten zu entdecken. In der ersten Phase des Projektes werden die tRNS Substrate des TET2 Enzyms identifiziert. In der zweiten Phase, werden der Einfluss von TET2-vermittelter Oxidation auf tRNS-Stabilität, die Tertiärstruktur, und Aminosäure-Beladung geprüft. In der dritten Phase, wird der Effekt von TET2-modifizierter tRNS auf das neue Proteom untersucht. Diese Arbeit wird einen wichtigen Einblick in die molekulare Funktion des TET2 Enzyms geben und einen neuen Mechanismus der Translationsregulation erforschen, der für die Krebsbiologie relevant ist.

Direct link to Lay Summary Last update: 26.12.2020

Responsible applicant and co-applicants

Abstract

Ten-Eleven Translocation-2 (TET2) is one of the most frequently mutated genes in several hematopoietic malignancies. Its tumor-suppressor function was previously connected with the ability of the TET2 enzyme to oxidize 5-methylcytosine of DNA. Only recently it was shown that TET2 is also capable of oxidizing RNA in vitro and in vivo. It was demonstrated that TET2 oxidizes tRNAs in HEK and mouse embryonic stem cells, and addition of the TET2-oxidized tRNAs was shown to stimulate translation in vitro by an unknown so far mechanism. My research goal is to determine the molecular mechanism underlying the enhanced translation by TET2-modified tRNAs and assess its involvement in the development of hematopoietic cancers. Specifically, with the help of the Swiss Early Postdoc Mobility Fellowship I will determine which tRNAs are oxidized by the TET2 enzyme, and at which positions, using several sequencing approaches. I will test if the TET2-mediated oxidation has an influence on the tertiary structure and stability of modified tRNAs using structural probing, northern blotting, and pulse-chase experiments. To address whether enhanced translation by the TET2-modified tRNAs is codon-specific, I will perform in vitro translation experiments using different reporter mRNAs. Investigation of the newly synthesized proteome under induced depletion of the TET2 enzyme combined with simultaneous transcription repression will allow me to analyze the direct effect of the tRNA oxidation on mammalian protein synthesis. Together, this work will provide a key insight into the molecular function of TET2 enzyme and might reveal a new mechanism of translation regulation relevant for cancer biology.
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