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April 11th, 2025
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The Ohio State University
biochemistry
biorxiv

An autism spectrum disorder mutation in Topoisomerase 3B causes accumulation of covalent mRNA intermediates by disrupting metal binding within the zinc finger domain

Warrick, J. E.Open in Google Scholar•Attili, D.Open in Google Scholar•van Eeuwen, T.Open in Google Scholar•Hoffmann-Weitsman, S. E.Open in Google Scholar•Forsyth, N. C.Open in Google Scholar•Barmada, S. J.Open in Google Scholar•Kearse, M. G.Open in Google Scholar

The loss and mutation of Topoisomerase 3{beta} (TOP3B), the only known eukaryotic topoisomerase with the ability to catalyze RNA strand passage reactions, is linked to schizophrenia, autism, and intellectual disability. Uniquely, TOP3B primarily localizes to the cytoplasm and has been shown to regulate mRNA translation and stability of a subset of transcripts. Three neurological disease-linked de novo TOP3B point mutations outside of the active site have been identified but their impact on TOP3B activity in cells remains poorly understood. Upon establishing a new Neuro2A-cell based TOP3B activity assay, we provide genetic and biochemical evidence that the autism-linked C666R mutation causes accumulation of unresolved TOP3B-mRNA covalent intermediates by directly disrupting metal coordination via a unique D1C3-type metal binding motif within the zinc finger domain. Furthermore, we show that primary neurons are sensitive to high levels of TOP3B-mRNA covalent intermediates and that such adducts are capable of causing ribosome collisions. Together, these data identify a previously underappreciated role of the zinc finger domain and how non-active site disease-linked mutations affect TOP3B activity.

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