Splicing factors Sf3A2 and Prp31 have direct roles in mitotic chromosome segregation

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Abstract

Several studies have shown that RNAi-mediated depletion of splicing factors (SFs) results in mitotic abnormalities. However, it is currently unclear whether these abnormalities reflect defective splicing of specific pre-mRNAs or a direct role of the SFs in mitosis. Here we show that two highly conserved SFs, Sf3A2 and Prp31, are required for chromosome segregation in both Drosophila and human cells. Injections of anti-Sf3A2 and anti-Prp31 antibodies into Drosophila embryos disrupt mitotic division within 1 minute, arguing strongly against a splicing-related mitotic function of these factors. We demonstrate that both SFs bind spindle microtubules (MTs) and the Ndc80 complex, which in Sf3A2- and Prp31-depleted cells is not tightly associated with the kinetochores; in HeLa cells the Ndc80/HEC1-SF interaction is restricted to the M phase. These results indicate that Sf3A2 and Prp31 directly regulate interactions among kinetochores, spindle microtubules and the Ndc80 complex in both Drosophila and human cells.

Data availability

All data generated or analyzed during this study are included in the manuscript and Supplementary File 1. The full dataset for proteomic analyses reported in Figures 6 and Supplementary File 1 can be found at https://www.thewakefieldlab.com/ms; the significantly reduced protein IDs for each RNAi experiment were interrogated using a Gene Ontology (GO) classifier (GOTermMapper (https://go.princeton.edu/cgi-bin/GOTermMapper), concentrating on the GO terms ""cell division"" (GO:0051301), ""mitotic cell cycle"" (GO:0000278) and ""chromosome segregation"" (GO:0007059). Source data files have been provided for Figures 1, 4, 5, 7, 8, and 9.

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Claudia Pellacani

Department of Biology and Biotechnology Charles Darwin, University of Rome, Rome, Italy

Competing interests
The authors declare that no competing interests exist.
Elisabetta Bucciarelli

Institute of Molecular Biology and Pathology, University of Rome, Rome, Italy

Competing interests
The authors declare that no competing interests exist.
Fioranna Renda

Department of Biology and Biotechnology Charles Darwin, University of Rome, Rome, Italy

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The authors declare that no competing interests exist.
Daniel Hayward

Biosciences/Living Systems Institute, University of Exeter, Exeter, United Kingdom

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The authors declare that no competing interests exist.
Antonella Palena

Institute of Molecular Biology and Pathology, University of Rome, Rome, Italy

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The authors declare that no competing interests exist.
Jack Chen

Biosciences/Living Systems Institute, University of Exeter, Exeter, United Kingdom

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The authors declare that no competing interests exist.
Silvia Bonaccorsi

Department of Biology and Biotechnology Charles Darwin, University of Rome, Roma, Italy

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The authors declare that no competing interests exist.
James G Wakefield

Biosciences/Living Systems Institute, University of Exeter, Exeter, United Kingdom

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The authors declare that no competing interests exist.
Maurizio Gatti

Department of Biology and Biotechnology Charles Darwin, University of Rome, Rome, Italy

For correspondence
maurizio.gatti@uniroma1.it

Competing interests
The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0003-3777-300X
Maria Patrizia Somma

Institute of Molecular Biology and Pathology, University of Rome, Rome, Italy

For correspondence
patrizia.somma@uniroma1.it

Competing interests
The authors declare that no competing interests exist.

Funding

Associazione Italiana per la Ricerca sul Cancro (IG16020)

Maurizio Gatti

Ministero dell'Istruzione, dell'Università e della Ricerca

Silvia Bonaccorsi

Biotechnology and Biological Sciences Research Council (BB/K017837/1)

James G Wakefield

Associazione Italiana per la Ricerca sul Cancro (IG20528)

Maurizio Gatti

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Copyright

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.