HITS-CLIP analysis of human ALKBH8 reveals interactions with fully processed substrate tRNAs and with specific noncoding RNAs

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This publication doesn't include Faculty of Economics and Administration. It includes Central European Institute of Technology. Official publication website can be found on muni.cz.
Authors

CAVALLIN Ivana BARTOŠOVIČ Marek SKALICKÝ Tomáš RENGARAJ Praveenkumar DEMKO Martin SCHMIDT-DENGLER Martina Christina DRINO Aleksej HELM Mark VAŇÁČOVÁ Štěpánka

Year of publication 2022
Type Article in Periodical
Magazine / Source RNA
MU Faculty or unit

Central European Institute of Technology

Citation
Web https://rnajournal.cshlp.org/content/28/12/1568
Doi http://dx.doi.org/10.1261/rna.079421.122
Keywords ALKBH8; Trm9; wobble uridine modification; mcm(5)U; mcm(5)s(2)U; HITS-CLIP
Attached files
Description Transfer RNAs acquire a large plethora of chemical modifications. Among those, modifications of the anticodon loop play important roles in translational fidelity and tRNA stability. Four human wobble U-containing tRNAs obtain 5-methoxycarbonylmethyluridine (mcm(5)U(34)) or 5-methoxycarbonylmethyl-2-thiouridine (mcm(5)s(2)U(34)), which play a role in decoding. This mark involves a cascade of enzymatic activities. The last step is mediated by alkylation repair homolog 8 (ALKBH8). In this study, we performed a transcriptome-wide analysis of the repertoire of ALKBH8 RNA targets. Using a combination of HITS-CLIP and RIP-seq analyses, we uncover ALKBH8-bound RNAs. We show that ALKBH8 targets fully processed and CCA modified tRNAs. Our analyses uncovered the previously known set of wobble U-containing tRNAs. In addition, both our approaches revealed ALKBH8 binding to several other types of noncoding RNAs, in particular C/D box snoRNAs.
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