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Grünberger, Felix ; Jüttner, Michael ; Knüppel, Robert Michael ; Ferreira-Cerca, Sébastien ; Grohmann, Dina

Nanopore-based RNA sequencing deciphers the formation, processing, and modification steps of rRNA intermediates in archaea

Grünberger, Felix , Jüttner, Michael, Knüppel, Robert Michael, Ferreira-Cerca, Sébastien und Grohmann, Dina (2023) Nanopore-based RNA sequencing deciphers the formation, processing, and modification steps of rRNA intermediates in archaea. RNA 29 (8), S. 1255-1273.

Veröffentlichungsdatum dieses Volltextes: 02 Nov 2023 16:20
Artikel
DOI zum Zitieren dieses Dokuments: 10.5283/epub.54952


Zusammenfassung

Ribosomal RNA (rRNA) maturation in archaea is a complex multistep process that requires well-defined endo- and exoribonuclease activities to generate fully mature linear rRNAs. However, technical challenges prevented detailed mapping of rRNA processing steps and a systematic analysis of rRNA maturation pathways across the tree of life. In this study, we used long-read (PCR)-cDNA and direct RNA ...

Ribosomal RNA (rRNA) maturation in archaea is a complex multistep process that requires well-defined endo- and exoribonuclease activities to generate fully mature linear rRNAs. However, technical challenges prevented detailed mapping of rRNA processing steps and a systematic analysis of rRNA maturation pathways across the tree of life. In this study, we used long-read (PCR)-cDNA and direct RNA nanopore-based sequencing to study rRNA maturation in three archaeal model organisms, namely the Euryarchaea Haloferax volcanii and Pyrococcus furiosus and the Crenarchaeon Sulfolobus acidocaldarius. Compared to standard short-read protocols, nanopore sequencing facilitates simultaneous readout of 5 & PRIME;- and 3 & PRIME;-positions, which is required for the classification of rRNA processing intermediates. More specifically, we (i) accurately detect and describe rRNA maturation stages by analysis of terminal read positions of cDNA reads and thereupon (ii) explore the stage-dependent installation of the KsgA-mediated dimethylations in H. volcanii using base-calling and signal characteristics of direct RNA reads. Due to the single-molecule sequencing capacity of nanopore sequencing, we could detect hitherto unknown intermediates with high confidence, revealing details about the maturation of archaea-specific circular rRNA intermediates. Taken together, our study delineates common principles and unique features of rRNA processing in euryarchaeal and crenarchaeal representatives, thereby significantly expanding our understanding of rRNA maturation pathways in archaea.



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Details

DokumentenartArtikel
Titel eines Journals oder einer ZeitschriftRNA
Verlag:COLD SPRING HARBOR LAB PRESS, PUBLICATIONS DEPT
Ort der Veröffentlichung:COLD SPRING HARBOR
Band:29
Nummer des Zeitschriftenheftes oder des Kapitels:8
Seitenbereich:S. 1255-1273
Datum16 Mai 2023
InstitutionenBiologie und Vorklinische Medizin > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Biochemie III
Biologie und Vorklinische Medizin > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Mikrobiologie (Archaeenzentrum) > Prof. Dr. Dina Grohmann
Identifikationsnummer
WertTyp
10.1261/rna.079636.123DOI
Stichwörter / KeywordsSULFOLOBUS-ACIDOCALDARIUS; QUALITY-CONTROL; CIRCULAR RNAS; INSIGHTS; 16S; ENDONUCLEASE; BIOGENESIS; SUBSTRATE; PRECURSOR; ROLES; Nanopore sequencing; ribosomal RNA; RNA modifications; RNA processing; archaea
Dewey-Dezimal-Klassifikation500 Naturwissenschaften und Mathematik > 540 Chemie
500 Naturwissenschaften und Mathematik > 570 Biowissenschaften, Biologie
StatusVeröffentlicht
BegutachtetJa, diese Version wurde begutachtet
An der Universität Regensburg entstandenJa
URN der UB Regensburgurn:nbn:de:bvb:355-epub-549520
Dokumenten-ID54952

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