TREX-2-ORC complex of D. melanogaster participates in nuclear export of histone MRNA

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Abstract

The TREX-2-ORC protein complex of D. melanogaster is necessary for the export of the bulk of synthesized poly(A)-containing mRNA molecules from the nucleus to the cytoplasm through the nuclear pores. However, the role of this complex in the export of other types of RNA remains unknown. We have shown that TREX-2-ORC participates in the nuclear export of histone mRNAs: it associates with histone mRNPs, binds to histone H3 mRNA at the 3’-terminal part of the coding region, and participates in the export of histone mRNAs from the nucleus to the cytoplasm.

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About the authors

M. M. Kurshakova

Engelhardt Institute of Molecular Biology RAS

Author for correspondence.
Email: kursha@mail.ru
Russian Federation, Moscow

Y. A. Yakusheva

Engelhardt Institute of Molecular Biology RAS

Email: kursha@mail.ru
Russian Federation, Moscow

S. G. Georgieva

Engelhardt Institute of Molecular Biology RAS

Email: kursha@mail.ru

Academician of the RAS

Russian Federation, Moscow

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Supplementary files

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2. Fig.1. TREX-2-ORC is associated with histone mRNA. (a) The results of immunosuppression of mRNP particles of the H2A, H3 and β-tubulin genes with antibodies to TREX-2-ORC subunits, IgG was used as a negative control. The results are presented as a percentage of the source material. All data are presented as an average ± standard deviation, and the Student's t-test was used to compare the control and the experiment. (*) indicates statistical significance with p-value <0.05, (**) indicates statistical significance with p-value < 0.01. (b) Western blot analysis of the deposition of H3 mRNP particles with a biotinylated antisense RNA probe to histone H3 mRNA using antibodies to Xmas-2, PCID2, ENY2, Orc3, Orc6, CPSF100, CPSF73. In the negative control (K-), an RNA probe with an antisense sequence to the RNA of the YFP protein was used. (c) Western blot analysis of the co-deposition of proteins with biotinylated semantic RNA probes to histone H3 mRNA sequences: full-length (H3), CDS1, CDS2, 3'UTR, using

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3. Fig. 2. The knockdown effect of TREX-2-ORC subunits on histone mRNA export. (a) Western blot analysis of protein expression levels in Xmas-2, Orc3 RNA interferences in S2 cells using appropriate antibodies. Staining with antibodies to lamin Dm0 was used as an equalizing control. (b) The results of the mRNA deposition of histone particles during Xmas-2 RNA interference with antibodies to TREX-2- ORC subunits were used, IgG was used as a negative control. The results are presented as a percentage of the source material. All data are presented as an average ± standard deviation, and the Student's t-test was used to compare control and experiment. (*) indicates the statistical significance of c p-value < 0.05, (**) indicates the statistical significance of c p-value<0.01. (c) Western blot analysis of RNA interferences Xmas-2, Orc3, NXF1 using antibodies to the same proteins. Staining with antibodies to lamin Dm0 was used as a leveling control. (d) FISH hybrid

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