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Using comparative transcriptomics we identified a small RNA, SQ528, that accumulates at higher levels in a CD25890 deletion mutant and we show that purified CD25890 cleaves SQ528 in a metal-dependent manner. Moreover, the overexpression of SQ528 increases sporulation under certain nutritional conditions phenocopying a CD25890 deletion mutant. CD25890 is an hexamer in solution and in vivo. An N-terminal domain, which self-interacts as assessed by size\u00a0exclusion chromatography and a two hybrid assay, is essential for oligomerization of CD25890. A C-terminal domain harbours residues H230, E254, and E258, conserved among orthologues, important for catalysis. AlphaFold2 modelling and cryo-EM suggest an elongated barrel-like structure with an internal cavity lined with basic residues that may aid in RNA binding. We show that CD25890 forms a complex with polynucleotide phosphorylase\u00a0which combines the endoribonuclease activity of the first with the exonucleolytic activity of the latter and leads to the complete degradation of SQ528. This study identifies a native substrate for the YicC-family of ribonucleases and advances our understanding of the role of CD25890 in sporulation initiation in C. difficile.<\/jats:p>","DOI":"10.1093\/nar\/gkaf644","type":"journal-article","created":{"date-parts":[[2025,7,12]],"date-time":"2025-07-12T16:39:52Z","timestamp":1752338392000},"source":"Crossref","is-referenced-by-count":2,"title":["An endoribonuclease of the YicC-like family delays sporulation via sRNA degradation in <i>Clostridioides difficile<\/i>"],"prefix":"10.1093","volume":"53","author":[{"given":"Diogo","family":"Martins","sequence":"first","affiliation":[{"name":"Instituto de Tecnologia Qu\u00edmica e Biol\u00f3gica Ant\u00f3nio Xavier, Universidade Nova de Lisboa , Avenida da Rep\u00fablica EAN , 2780-157 Oeiras ,","place":["Portugal"]}]},{"given":"Bruno","family":"Salgueiro","sequence":"additional","affiliation":[{"name":"Instituto de Tecnologia Qu\u00edmica e Biol\u00f3gica Ant\u00f3nio Xavier, Universidade Nova de Lisboa , Avenida da Rep\u00fablica EAN , 2780-157 Oeiras ,","place":["Portugal"]}]},{"given":"Daniel","family":"Sobral","sequence":"additional","affiliation":[{"name":"Genomics and Bioinformatics Unit , Department of Infectious Diseases, National Institute of Health Doutor Ricardo Jorge (INSA), Av. Padre Cruz, 1649-016 Lisboa ,","place":["Portugal"]}]},{"given":"Marcos","family":"Gragera","sequence":"additional","affiliation":[{"name":"Centro Nacional de Biotecnolog\u00eda (CNB), CSIC , C. Darwin, 3, Fuencarral-El Pardo, 28049 Madrid,","place":["Spain"]}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4348-6229","authenticated-orcid":false,"given":"Zach","family":"Hensel","sequence":"additional","affiliation":[{"name":"Instituto de Tecnologia Qu\u00edmica e Biol\u00f3gica Ant\u00f3nio Xavier, Universidade Nova de Lisboa , Avenida da Rep\u00fablica EAN , 2780-157 Oeiras ,","place":["Portugal"]}]},{"given":"Adriano\u00a0O","family":"Henriques","sequence":"additional","affiliation":[{"name":"Instituto de Tecnologia Qu\u00edmica e Biol\u00f3gica Ant\u00f3nio Xavier, Universidade Nova de Lisboa , Avenida da Rep\u00fablica EAN , 2780-157 Oeiras ,","place":["Portugal"]}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9474-2405","authenticated-orcid":false,"given":"C\u00e9lia\u00a0V","family":"Rom\u00e3o","sequence":"additional","affiliation":[{"name":"Instituto de Tecnologia Qu\u00edmica e Biol\u00f3gica Ant\u00f3nio Xavier, Universidade Nova de Lisboa , Avenida da Rep\u00fablica EAN , 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