Evolutionarily conserved DHX9/MLE helicase is involved in the regulation of its own mRNA expression level in Drosophila melanogaster

Cover Page

Cite item

Full Text

Open Access Open Access
Restricted Access Access granted
Restricted Access Subscription Access

Abstract

The MLE helicase of D. melanogaster, like its ortholog DHX9 in mammals, is involved in a wide range of processes related to the regulation of gene expression. In the present study, we investigated the impact of the mle[9] mutation on its own mRNA expression level. It was shown that in addition to the previously described deletion in the catalytic domain of the protein, which impairs its helicase activity, the mle[9] mutation contains an additional small deletion in the C-terminal domain. In the mle[9] mutation background, there was a threefold increase in the expression of the main transcript of the mle gene encoding the full-length protein. Binding of MLE to chromatin at the coding region and promoters of the mle gene and nearby enhancers was analyzed. To exclude the influence of dosage compensation, experiments were performed on females. The data obtained indicate the role of MLE in specific regulation of its own mRNA expression level in vivo at the adult stage.

Full Text

Restricted Access

About the authors

I. A. Zolin

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

Email: julia.v.nikolenko@gmail.com
Russian Federation, Moscow

S. G. Georgieva

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

Email: julia.v.nikolenko@gmail.com

Academician of the RAS

Russian Federation, Moscow

J. V. Nikolenko

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

Author for correspondence.
Email: julia.v.nikolenko@gmail.com
Russian Federation, Moscow

References

  1. Lee T., Pelletier J. The biology of DHX9 and its potential as a therapeutic target. Oncotarget. 2016. Vol. 7. P. 42716–42739.
  2. Gulliver C., Hoffmann R., Baillie G.S. The enigmatic helicase DHX9 and its association with the hallmarks of cancer. Future Sci OA. 2020. Vol. 7. FSO650.
  3. Kuroda M.I., Kernan M.J., Kreber R., et al. The maleless protein associates with the X chromosome to regulate dosage compensation in drosophila. Cell. 1991. Vol. 66. P. 935–947.
  4. Николенко Ю.В., Георгиева С.Г., Копытова Д.В. Разнообразие функций хеликазы MLE в регуляции экспрессии генов у высших эукариот. Молекулярная биология. 2023. Т. 57. С. 1–14.
  5. Николенко Ю.В., Куршакова М.М., Краснов А.Н. Мультифункциональный белок ENY2 взаимодействует с РНК-хеликазой MLE. Докл. Акад. Наук. 2019. Т. 489. С. 637–640.
  6. Николенко Ю.В., Куршакова М.М., Краснов А.Н., и др. Хеликаза MLE – новый участник регуляции транскрипции гена ftz-f1, кодирующего ядерный рецептор у высших эукариот. Докл. Акад. Наук. Науки о жизни. 2021. Т. 496. С.48–51.
  7. Kotlikova I. V, Demakova O. V, Semeshin V.F., et al. The Drosophila Dosage Compensation Complex Binds to Polytene Chromosomes Independently of Developmental Changes in Transcription. Genetics. 2006. Vol. 14. P. 1478–1488.
  8. Cugusi S., Li Y., Jin P., et al. The Drosophila Helicase MLE Targets Hairpin Structures in Genomic Transcripts. PLoS Genet. 2016. Vol. 12. P. e1005761.
  9. Cugusi S., Kallappagoudar S., Ling H., et al. The Drosophila Helicase Maleless (MLE) is Implicated in Functions Distinct From its Role in Dosage Compensation. Molecular & Cellular Proteomics. 2015. Vol. 14. P. 1478–1488.
  10. Kernan M.J., Kuroda M.I., Kreber R., et al. napts, a Mutation affecting sodium channel activity in Drosophila, Is an allele of mle a regulator of X chromosome transcription. Cell. 1991. Vol. 66. P. 949–959.
  11. Ашниев Г.А., Георгиева С.Г., Николенко Ю.В. Функции хеликазы MLE Drosophila melanogaster вне дозовой компенсации: молекулярная природа и плейотропный эффект мутации mle[9]. Генетика. 2024. Т. 60. С.34–46.
  12. Izzo A., Regnard C., Morales V., et al. Structure-function analysis of the RNA helicase maleless. Nucleic Acids Res. 2008. Vol. 36. P. 950–962.
  13. Kopytova D.V., Krasnov A.N., Orlova A.V., et al. ENY2: Couple, triple...more? Cell Cycle. 2010. Vol. 9. P. 479–481.
  14. Николенко Ю.В., Краснов А.Н., Мазина М.Ю., и др. Изучение свойств нового экдизонзависимого энхансера. Докл. Акад. Наук. 2017. Т. 474. С. 756–759.
  15. Arnold C.D., Gerlach D., Stelzer C., et al. Genome-wide quantitative enhancer activity maps identified by STARR-seq. Science. 2013. Vol. 339. P. 1074–1077.
  16. Reenan R.A., Hanrahan C.J., Ganetzky B. The mlenapts RNA Helicase Mutation in Drosophila Results in a Splicing Catastrophe of the para Na + Channel Transcript in a Region of RNA Editing. Neuron. 2000. Vol. 25. P. 139–149.
  17. Aratani S., Fujii R., Fujita H., et al. Aromatic residues are required for RNA helicase A mediated transactivation. Int J Mol Med. 2003. Vol. 12. P. 175–180.
  18. Aratani S., Kageyama Y., Nakamura A., et al. MLE activates transcription via the minimal transactivation domain in Drosophila. Int J Mol Med. 2008. Vol. 21. P. 469–476.
  19. Calame D.G. et al. Monoallelic variation in DHX9, the gene encoding the DExH-box helicase DHX9, underlies neurodevelopment disorders and Charcot-Marie-Tooth disease. Am J Hum Genet. 2023. Vol. 110. P. 1394–1413.
  20. Fergestad T., Ganetzky B., Palladino M.J. Neuropathology in Drosophila membrane excitability mutants. Genetics. 2006. Vol. 172. P. 1031–1042.

Supplementary files

Supplementary Files
Action
1. JATS XML
Download
2. Fig. 1. The mle gene and the domain structure of the MLE protein under normal conditions and against the background of the mle[9] mutation (quoted from [11] with modifications). a – schematic representation of the mle gene and its transcripts: mle-RA is the transcript encoding the full-length protein, mle-RC is the alternative transcript. Exons are indicated by rectangles, the coding region is highlighted in gray. Arrows 1, 2, 3 indicate the position of the primers used to measure the transcription level of mle-RA and mle-RC. Arrows above the gene indicate the position of deletions in the mle[9] mutation: gray – the previously described deletion in the catalytic “core”, black – the deletion in the C-terminal domain discovered in the present study. Asterisks indicate the position of the nearby enhancers STARR_OSC-4142 and STARR_OSC-2528. b – domain structure of the MLE protein. Arrows indicate the positions of deletions in the mle[9] mutation: gray – previously described deletion in the catalytic core, black – 7 aa deletion in the C-terminal domain discovered in the present study. Regions corresponding to specific domains are shown in gray, linker regions are shown in white. Shown are two dsRNA-binding domains – dsRBD1 and ds-RBD2, minimal transactivation domain MTAD, catalytic core consisting of RecA1 and RecA2 domains and helicase-associated domain HA2, oligosaccharide/oligonucleotide binding region OB-fold, C-terminal domain RGG-box including nuclear localization signal NLS and glycine-rich sequence Gly. Conserved motifs in the catalytic core are indicated by Roman numerals.

Download (87KB)
Indexing metadata
3. Fig. 2. Study of the effect of MLE on the expression of its own gene in vivo in D. melanogaster females at the imago stage. a – changes in the expression level of the mle gene transcripts against the background of the mle mutation[9]. The level of mle transcripts was measured relative to the level of the control gene Rpl32 transcript. The mle-RA value in wild-type females in one of three experimental replicates was taken as 1 on the ordinate axis. * – p<0.01. b – analysis of binding of the wild-type MLE protein (mle+) and the mutant protein (mle[9]) to the promoters of their own gene, to coding regions in the 5’-region of the gene (+460 – distance in bp from the mle-RC transcription start site) and in the 3’-region of the gene (+4470 – distance in bp from the mle-RC transcription start site) and to nearby enhancers (STARR_OSC-4142 and STARR_OSC-2528).

Download (129KB)
Indexing metadata

Copyright (c) 2025 Russian Academy of Sciences