PXD067005

PXD067005 is an original dataset announced via ProteomeXchange.

Title	A systematic screen of archaeal MazF proteins reveals Tth-MazF1, a versatile, sequence-specific ribonuclease from Thermococcus thioreducens.
Description	Nucleotide- and sequence-specific ribonucleases are essential tools for dissecting RNA structure, modification, and composition, yet the enzymatic repertoire available for high-resolution RNA analysis remains limited. Here, we systematically screened 186 MazF homologs, including 164 from archaeal organisms, and identified 132 active endoribonucleases with diverse sequence preferences. Using a semi-high-throughput expression and purification workflow combined with an Illumina-based RNA-seq assay, we defined recognition motifs for 90 enzymes, uncovering a striking diversity of UG-rich cleavage specificities across the archaeal domain. Among these, Tth-MazF1, a thermostable endoribonuclease from the hyperthermophilic archaeon Thermococcus thioreducens, displayed robust and specific cleavage at U↓GN motifs across a broad range of temperatures (4 ˚C to 85 ˚C). Tth-MazF1 enabled high sequence coverage of 0.8-8.9 kb mRNA transcripts by LC-MS/MS and direct detection of key structural features, including the 5′ cap, poly(A) tail, and mass-altering epitranscriptomic modifications in rRNA. Tolerance of Tth-MazF1 to high temperatures, salt, and chemical modifications highlights it as a powerful enzymatic tool for analyzing chemically modified and structured RNAs. These findings position archaeal MazF proteins as a rich source of customizable RNases for biotechnology applications, such as mass spectrometry-based analysis of RNA-based vaccines and therapeutics as well as cellular RNAs.
HostingRepository	PRIDE
AnnounceDate	2026-05-25
AnnouncementXML	Submission_2026-05-24_16:44:06.823.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Eric Wolf
SpeciesList	scientific name: Thermococcus thioreducens; NCBI TaxID: NEWT:277988;
ModificationList	No PTMs are included in the dataset
Instrument	Orbitrap Eclipse

Revision	Datetime	Status	ChangeLog Entry
0	2025-08-06 10:19:09	ID requested
⏵ 1	2026-05-24 16:44:07	announced

Jepson BF, Wolf EJ, Dawson AR, Jin J, Potapov V, Dai N, Corr, ê, a IR, Yigit E, Systematic screening of archaeal MazF homologs reveals Tth-MazF1, a versatile, sequence-specific ribonuclease from Thermococcus thioreducens. Nucleic Acids Res, 54(8):(2026) [pubmed]

10.1093/nar/gkag338;

submitter keyword: Thermococcus thioreducens RNA LC-MS/MS

Ivan R. Correa Jr.
contact affiliation	New England Biolabs, RNA Division
contact email	correa@neb.com
lab head
Eric Wolf
contact affiliation	New England Biolabs
contact email	ewolf@neb.com
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD067005
     1. Label: PRIDE project
     2. Name: A systematic screen of archaeal MazF proteins reveals Tth-MazF1, a versatile, sequence-specific ribonuclease from Thermococcus thioreducens.