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bioRxiv · 10.1101/2025.02.21.638496

Involvement of RNase J in CRISPR RNA maturation and degradosome formation in cyanobacteria

Bilger, R.·Drepper, F.·Knapp, B.·Berndt, T.·Landerer, H.·Putzer, H.·Huesgen, P.·Hess, W. R.
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Abstract

Many bacteria and archaea use CRISPR-Cas systems, which provide RNA-based, adaptive, and inheritable immune defenses against invading viruses and other foreign genetic elements. The proper processing of CRISPR guide RNAs (crRNAs) is a crucial step in the maturation of the defense complexes and is frequently performed by specialized ribonucleases encoded by cas genes. However, some systems employ enzymes associated with degradosome or housekeeping functions, such as RNase III or the endoribonuclease RNase E. Here, the endo- and 5'-exoribonuclease RNase J was identified as an additional enzyme involved in crRNA maturation, acting jointly with RNase E in the crRNA maturation of a type III-Bv CRISPR-Cas system, and possibly together with a further RNase in the cyanobacterium Synechocystis sp. PCC 6803. Co-IP experiments revealed a small set of proteins that were co-enriched with RNase J, among them the exoribonuclease polyribonucleotide nucleotidyltransferase (PNPase). Despite a measured, strong 3 exonucleolytic activity of the recombinant enzyme, PNPase was not confirmed to contribute to crRNA maturation. However, the co-IP results indicate that PNPase in Synechocystis is an enzyme that can recruit either RNase E or RNase J, together with additional proteins.

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Bilger, R., Drepper, F., Knapp, B., Berndt, T., Landerer, H., Putzer, H., Huesgen, P., Hess, W. R.. 2025-02-23. Involvement of RNase J in CRISPR RNA maturation and degradosome formation in cyanobacteria. https://doi.org/10.1101/2025.02.21.638496

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