Detection and quantification of glycosylated queuosine modified tRNAs by acid denaturing and APB gels.

ABSTRACT: Queuosine (Q) is a conserved tRNA modification in bacteria and eukaryotes. Eukaryotic Q-tRNA modification occurs through replacing the guanine base with the scavenged metabolite queuine at the wobble position of tRNAs with G ₃₄U₃₅N₃₆ anticodon (Tyr, His, Asn, Asp) by the QTRT1/QTRT2 heterodimeric enzyme encoded in the genome. In humans, Q-modification in tRNA^Tyr and tRNA^Asp are further glycosylated with galactose and mannose, respectively. Although galactosyl-Q (galQ) and mannosyl-Q (manQ) can be measured by LC/MS approaches, the difficulty of detecting and quantifying these modifications with low sample inputs has hindered their biological investigations. Here we describe a simple acid denaturing gel and nonradioactive northern

SUBMITTER: Zhang W 

PROVIDER: S-EPMC7430669 | biostudies-literature | 2020 Sep

REPOSITORIES: biostudies-literature