{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,2]],"date-time":"2026-04-02T05:50:29Z","timestamp":1775109029041,"version":"3.50.1"},"reference-count":43,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2026,3,31]],"date-time":"2026-03-31T00:00:00Z","timestamp":1774915200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100005188","name":"California Tobacco Related Diseases Research Program","doi-asserted-by":"crossref","award":["#T30DT0811"],"award-info":[{"award-number":["#T30DT0811"]}],"id":[{"id":"10.13039\/100005188","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000054","name":"NCI","doi-asserted-by":"publisher","award":["R37CA292795"],"award-info":[{"award-number":["R37CA292795"]}],"id":[{"id":"10.13039\/100000054","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["ncRNA"],"abstract":"<jats:p>Background\/Objectives: Canonical microRNAs possess a 5\u2032 phosphate required for Argonaute binding and activity. However, prior work identified an unphosphorylated, inactive nuclear pool of the important radiation-responsive microRNA, miR-34, that is rapidly phosphorylated and activated in response to ionizing radiation (IR). Here, we extend this work and investigate the role of paraspeckles, a phase-separated nuclear sub-compartment, and their association with the localization of unphosphorylated miR-34a. Methods: Mass spectrometry was performed to identify interacting partners of unphosphorylated mir-34. CRISPR-mediated deletion of the paraspeckle NEAT1_2 triple helix motif was performed to create an A549 cell line lacking paraspeckles (dTH). Activity and expression of mir-34a post-irradiation were evaluated by qRT-PCR and luciferase assays comparing dTH and wild-type (WT) A549 cell lines. In situ hybridization (ISH) was performed to evaluate mir-34a localization before and after IR, comparing dTH and WT cell lines. Results: Mass spectrometry identified paraspeckle proteins as significantly enriched interacting partners of unphosphorylated mir-34 mimics. By qRT-PCR and luciferase assays, we found that paraspeckle loss prevented radiation-induced early activation of unphosphorylated mir-34a. We found no difference in radiation-induced transcription of pri-miR-34a, but early processing to pre-miR-34a appeared delayed. ISH confirmed that loss of paraspeckles altered the nuclear localization of miR-34a before and after IR. Conclusions: These data suggest that paraspeckles are associated with nuclear localization and early radiation-responsive activation of unphosphorylated miR-34a. This suggests a coordinated nuclear sequestration of this important miR in its unphosphorylated state to enable an enhanced radiation response.<\/jats:p>","DOI":"10.3390\/ncrna12020012","type":"journal-article","created":{"date-parts":[[2026,3,31]],"date-time":"2026-03-31T16:34:08Z","timestamp":1774974848000},"page":"12","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Paraspeckles Are Associated with the Activation and Nuclear Localization of Unphosphorylated miR-34a"],"prefix":"10.3390","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7716-6506","authenticated-orcid":false,"given":"Graham H.","family":"Read","sequence":"first","affiliation":[{"name":"Radiation Oncology Department, University of California, Los Angeles, CA 90095, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kristen","family":"McGreevy","sequence":"additional","affiliation":[{"name":"Biostatistics, MiraDx, Los Angeles, CA 90025, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-5647-0565","authenticated-orcid":false,"given":"Hanny","family":"Issawi","sequence":"additional","affiliation":[{"name":"Radiation Oncology Department, University of California, Los Angeles, CA 90095, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tiffany","family":"Yang","sequence":"additional","affiliation":[{"name":"Radiation Oncology Department, University of California, Los Angeles, CA 90095, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Cynthia","family":"Tsang","sequence":"additional","affiliation":[{"name":"Radiation Oncology Department, University of California, Los Angeles, CA 90095, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ihsan A.","family":"Turk","sequence":"additional","affiliation":[{"name":"Radiation Oncology Department, University of California, Los Angeles, CA 90095, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3798-0922","authenticated-orcid":false,"given":"Emily","family":"Rietdorf","sequence":"additional","affiliation":[{"name":"Radiation Oncology Department, University of California, Los Angeles, CA 90095, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Whitaker","family":"Cohn","sequence":"additional","affiliation":[{"name":"Department of Biostatistics, University of California, Los Angeles, CA 90095, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"David W.","family":"Salzman","sequence":"additional","affiliation":[{"name":"Radiation Oncology Department, University of California, Los Angeles, CA 90095, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2763-7733","authenticated-orcid":false,"given":"Julian P.","family":"Whitelegge","sequence":"additional","affiliation":[{"name":"Pasarow Mass Spectrometry Lab, University of California, Los Angeles, CA 90095, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5096-3281","authenticated-orcid":false,"given":"Joanne B.","family":"Weidhaas","sequence":"additional","affiliation":[{"name":"Radiation Oncology Department, University of California, Los Angeles, CA 90095, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2026,3,31]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"597","DOI":"10.1038\/nrg2843","article-title":"The widespread regulation of microRNA biogenesis, function and decay","volume":"11","author":"Krol","year":"2010","journal-title":"Nat. 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