Global analysis by LC-MS/MS of N6-methyladenosine and inosine in mRNA reveal complex incidence

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388963%3A_____%2F25%3A00618830" target="_blank" >RIV/61388963:_____/25:00618830 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/00216224:14740/25:00141605

Výsledek na webu

<a href="http://www.rnajournal.org/cgi/doi/10.1261/rna.080324.124" target="_blank" >http://www.rnajournal.org/cgi/doi/10.1261/rna.080324.124</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1261/rna.080324.124" target="_blank" >10.1261/rna.080324.124</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Global analysis by LC-MS/MS of N6-methyladenosine and inosine in mRNA reveal complex incidence

Popis výsledku v původním jazyce

The precise and unambiguous detection and quantification of internal RNA modifications represents a critical step for understanding their physiological functions. The methods of direct RNA sequencing are quickly developing allowing for the precise location of internal RNA marks. This detection is, however, not quantitative and still presents detection limits. One of the biggest remaining challenges in the field is still the detection and quantification of m6A, m6Am, inosine, and m1A modifications of adenosine. The second intriguing and timely question remaining to be addressed is the extent to which individual marks are coregulated or potentially can affect each other. Here, we present a methodological approach to detect and quantify several key mRNA modifications in human total RNA and in mRNA, which is difficult to purify away from contaminating tRNA. We show that the adenosine demethylase FTO primarily targets m6Am marks in noncoding RNAs in HEK293T cells. Surprisingly, we observe little effect of FTO or ALKBH5 depletion on the m6A mRNA levels. Interestingly, the upregulation of ALKBH5 is accompanied by an increase in inosine level in overall mRNA.

Název v anglickém jazyce

Global analysis by LC-MS/MS of N6-methyladenosine and inosine in mRNA reveal complex incidence

Popis výsledku anglicky

The precise and unambiguous detection and quantification of internal RNA modifications represents a critical step for understanding their physiological functions. The methods of direct RNA sequencing are quickly developing allowing for the precise location of internal RNA marks. This detection is, however, not quantitative and still presents detection limits. One of the biggest remaining challenges in the field is still the detection and quantification of m6A, m6Am, inosine, and m1A modifications of adenosine. The second intriguing and timely question remaining to be addressed is the extent to which individual marks are coregulated or potentially can affect each other. Here, we present a methodological approach to detect and quantify several key mRNA modifications in human total RNA and in mRNA, which is difficult to purify away from contaminating tRNA. We show that the adenosine demethylase FTO primarily targets m6Am marks in noncoding RNAs in HEK293T cells. Surprisingly, we observe little effect of FTO or ALKBH5 depletion on the m6A mRNA levels. Interestingly, the upregulation of ALKBH5 is accompanied by an increase in inosine level in overall mRNA.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10608 - Biochemistry and molecular biology

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2025

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

RNA

ISSN

1355-8382

e-ISSN

1469-9001

Svazek periodika

31

Číslo periodika v rámci svazku

4

Stát vydavatele periodika

US - Spojené státy americké

Počet stran výsledku

15

Strana od-do

514-528

Kód UT WoS článku

001620028500001

EID výsledku v databázi Scopus

2-s2.0-105000309599