TY - JOUR
T1 - Combinatorial control of Spo11 alternative splicing by modulation of RNA polymerase II dynamics and splicing factor recruitment during meiosis
AU - Cesari, Eleonora
AU - Loiarro, Maria
AU - Naro, Chiara
AU - Pieraccioli, Marco
AU - Farini, Donatella
AU - Pellegrini, Livia
AU - Pagliarini, Vittoria
AU - Bielli, Pamela
AU - Sette, Claudio
N1 - Funding Information:
We wish to thank Dr. M. Barchi for fruitful discussion throughout this work and Drs. M.A. Handel, B. Chabot, D. Elliott for providing antibodies used in this study, Dr. E. Buratti for providing the TDP-43 vector. This work was supported by Telethon (GGP12189), by Associazione Italiana Ricerca sul Cancro (AIRC) and by the UCSC grant Linea D1. C.N. was supported in part by a scholarship from Fondazione Umberto Veronesi. Supplementary information is available at Cell Death & Disease’s website.
Publisher Copyright:
© 2020, The Author(s).
Copyright:
Copyright 2020 Elsevier B.V., All rights reserved.
PY - 2020/4/1
Y1 - 2020/4/1
N2 - Homologous recombination and chromosome segregation in meiosis rely on the timely expression of two splice variants of the endonuclease SPO11, named α and β, which respectively skip or include exon 2. However, in spite of its physiological importance, the mechanism underlying Spo11 alternative splicing in meiosis is still unknown. By screening the activity of factors that are predicted to bind the alternatively spliced region of Spo11, we identified hnRNPH as a key regulator of SPO11α splicing in mouse spermatocytes. Although hnRNPH was not upregulated in meiosis concomitantly with the switch in splicing, its recruitment to Spo11 pre-mRNA was favored by selective modulation of RNA polymerase II (RNAPII) phosphorylation and processivity in proximity of exon 2. The hnRNPH binding sites were localized near those of splicing factors that promote SPO11β splicing, suggesting that hnRNPH favors exon 2 skipping by competing out positive regulators. Indeed, hnRNPH binds proximal to a consensus motif for Sam68, a positive regulator of SPO11β splicing in vitro and in vivo, and it interferes with Sam68 binding to the Spo11 pre-mRNA. Thus, our work reveals that modulation of RNAPII dynamics in concert with hnRNPH recruitment exerts a combinatorial control of the timely regulated Spo11 splicing during meiosis.
AB - Homologous recombination and chromosome segregation in meiosis rely on the timely expression of two splice variants of the endonuclease SPO11, named α and β, which respectively skip or include exon 2. However, in spite of its physiological importance, the mechanism underlying Spo11 alternative splicing in meiosis is still unknown. By screening the activity of factors that are predicted to bind the alternatively spliced region of Spo11, we identified hnRNPH as a key regulator of SPO11α splicing in mouse spermatocytes. Although hnRNPH was not upregulated in meiosis concomitantly with the switch in splicing, its recruitment to Spo11 pre-mRNA was favored by selective modulation of RNA polymerase II (RNAPII) phosphorylation and processivity in proximity of exon 2. The hnRNPH binding sites were localized near those of splicing factors that promote SPO11β splicing, suggesting that hnRNPH favors exon 2 skipping by competing out positive regulators. Indeed, hnRNPH binds proximal to a consensus motif for Sam68, a positive regulator of SPO11β splicing in vitro and in vivo, and it interferes with Sam68 binding to the Spo11 pre-mRNA. Thus, our work reveals that modulation of RNAPII dynamics in concert with hnRNPH recruitment exerts a combinatorial control of the timely regulated Spo11 splicing during meiosis.
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U2 - 10.1038/s41419-020-2443-y
DO - 10.1038/s41419-020-2443-y
M3 - Article
C2 - 32303676
AN - SCOPUS:85083786291
VL - 11
JO - Cell Death and Disease
JF - Cell Death and Disease
SN - 2041-4889
IS - 4
M1 - 240
ER -