Two RNA-binding regions of PCID2, subunit of the nuclear mRNA export complex TREX-2, competitively interact with the 3'-noncoding region of RAS2 mRNA

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Abstract

PCID2 protein is a subunit of the eukaryotic complex TREX-2, which is responsible for nuclear export of mRNA. PCID2 plays an important role in the complex, being responsible for the recognition and binding of the mRNA molecule. PCID2 of D. melanogaster interacts with a region of ras2 (fr4_2) mRNA and has two interaction sites located in its PCI domain: the M-PCID2 region, which non-specific binds to mRNA and the C-terminal part (C-PCID2), which specifically recognizes the ras2 fr4_2 mRNA sequence. At the same time, specific binding to C-PCID2 requires a preliminary nonspecific interaction with M-PCID2. It remains unclear how the transition from a primary nonspecific interaction to a specific interaction occurs. Whether both regions interact with RNA simultaneously or whether the nonspecific interaction is only necessary in the first step for subsequent specific binding. This study has shown that binding of M-PCID2 and C-PCID2 to ras2 fr4_2 RNA is competitive. M-PCID2 binds more efficiently and displaces C-PCID2 from the complex with the ras2 mRNA fragment. Thus, additional factors are required to replace the M-PCID2 contact by C-PCID2 during the interaction of full-length PCID2 protein with ras2 mRNA. We also show that point mutations in M-PCID2 that disrupt the interaction of full-length PCID2 with RNA result in greater association of M-PCID2 with RNA. It is likely that the increased affinity of M-PCID2 for RNA disrupts the ability to replace M-PCID2 with C-PCID2 within full-length PCID2.

About the authors

Y. A. Vdovina

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

Email: d_dmitrieva@mail.ru
Moscow, Russian Federation

S. G. Georgieva

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

Email: d_dmitrieva@mail.ru
Moscow, Russian Federation

D. V. Kopytova

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

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

References

  1. Lu Q. et al. Arabidopsis homolog of the yeast TREX-2 mRNA export complex: components and anchoring nucleoporin // The Plant journal: for cell and molecular biology. England, 2010. Vol. 61, № 2. P. 259–270.
  2. Luna R., Gonzalez-Aguilera C., Aguilera A. Transcription at the proximity of the nuclear pore: a role for the THP1-SAC3-SUS1-CDC31 (THSC) complex // RNA biology. United States, 2009. Vol. 6, № 2. P. 145–148.
  3. Rodríguez-Navarro S. et al. Sus1, a functional component of the SAGA histone acetylase complex and the nuclear pore-associated mRNA export machinery // Cell. United States, 2004. Vol. 116, № 1. P. 75–86.
  4. Fischer T. et al. The mRNA export machinery requires the novel Sac3p-Thp1p complex to dock at the nucleoplasmic entrance of the nuclear pores // The EMBO journal. England, 2002. Vol. 21, № 21. P. 5843–5852.
  5. Gonzalez-Aguilera C. et al. The THP1-SAC3-SUS1-CDC31 complex works in transcription elongation-mRNA export preventing RNA-mediated genome instability // Molecular biology of the cell. United States, 2008. Vol. 19, № 10. P. 4310–4318.
  6. Jani D. et al. Functional and structural characterization of the mammalian TREX-2 complex that links transcription with nuclear messenger RNA export // Nucleic acids research. England, 2012. Vol. 40, № 10. P. 4562–4573.
  7. Kopytova D. et al. ORC interacts with THSC/TREX-2 and its subunits promote Nxf1 association with mRNP and mRNA export in Drosophila // Nucleic Acids Research. 2016. Vol. 44, № 10. P. 4920–4933.
  8. Glukhova A. A. et al. PCID2, a subunit of the Drosophila TREX-2 nuclear export complex, is essential for both mRNA nuclear export and its subsequent cytoplasmic trafficking // RNA Biology. United States, 2021. Vol. 18, № 11. P. 1969–1980.
  9. Vdovina Y. A. et al. PCID2 Subunit of the Drosophila TREX-2 Complex Has Two RNA-Binding Regions // Current issues in molecular biology. Switzerland, 2023. Vol. 45, № 7. P. 5662–5676.
  10. Ellisdon A. M. et al. Structural basis for the assembly and nucleic acid binding of the TREX-2 transcription-export complex // Nature structural & molecular biology. United States, 2012. Vol. 19, № 3. P. 328–336.
  11. Stewart M. Structure and Function of the TREX-2 Complex // Sub-cellular biochemistry. United States, 2019. Vol. 93. P. 461–470.
  12. Вдовина Ю. А., Геогиева С. Г., Копытова Д. В. Точечные мутации в М-домене PCID2 нарушают его функции в экспорте мРНК у Drosophila melanogaster // Доклады Российской академии наук. Науки о жизни. 2024. Т. 518. С. 5–9.
  13. Golovanov A. P. et al. The solution structure of REF2-I reveals interdomain interactions and regions involved in binding mRNA export factors and RNA // RNA (New York, NY). 2006. Vol. 12, № 11. P. 1933–1948.
  14. Hautbergue G. M. et al. Mutually exclusive interactions drive handover of mRNA from export adaptors to TAP // Proceedings of the National Academy of Sciences of the United States of America. United States, 2008. Vol. 105, № 13. P. 5154–5159.

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