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The influence of novel transcriptional regulatory element in intron 14 on the expression of Janus kinase 2 gene in myeloproliferative neoplasms

  • Human Genetics • Original Paper
  • Published:
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Abstract

The expression of Janus kinase 2 (JAK2) gene is altered in myeloproliferative neoplasms (MPN) and the regulation of transcription could be a mechanism that modulates JAK2 gene expression. We analyzed the transcriptional potential of single-nucleotide polymorphism (SNP) rs12343867 T > C in JAK2 intron 14, tagging 46/1 haplotype, and its influence on JAK2 gene expression. Functional analysis of JAK2 intron 14 was performed using the pBLCAT5 reporter system in K562 cells. Identification of the proteins binding to the intron 14 regulatory element was accomplished by electrophoretic mobility shift assay (EMSA) and supershift assays. Quantification of the expression of JAK2 gene in a cohort of 51 MPN patients and 12 healthy controls was performed by real-time quantitative polymerase chain reaction (RQ-PCR). Functional analysis revealed that the intronic DNA element harboring SNP rs12343867 T > C acts as a transcriptional repressor in vitro. The repressor activity was significantly attenuated by the presence of nucleotide C. Supershift analysis showed the enrolment of transcriptional factor Meis1 in this process. RQ-PCR experiments showed increased JAK2 expression in patients with the JAK2V617F mutation, with a significant difference between essential thrombocythemia (ET), polycythemia vera (PV), and myelofibrosis (MF) patients. SNP rs12343867 showed no statistically significant influence on the expression of JAK2 gene in MNP patients.

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Acknowledgments

The authors would like to thank Dr. Milena Stevanovic for providing the TGIF and Meis1/2 antibodies. This work was supported by the Ministry of Education and Science, Republic of Serbia (grant no. III41004).

Authorship and disclosures

S.P. was the principal investigator and takes primary responsibility for the paper. S.P. and V.S. designed the study; M.C. performed the case and control sample collection and clinical management of patients; V.S. and M.R. performed the laboratory work; M.S., B.Z., and S.S. analyzed the data and participated in the drafting of the paper; T.K.-D. and N.T. performed the statistical analysis; V.S. and G.N. wrote the final version of the paper. The authors report no potential conflicts of interest.

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Authors and Affiliations

  1. Institute of Molecular Genetics and Genetic Engineering, University of Belgrade, Belgrade, Serbia

    Vesna Spasovski, Natasa Tosic, Gordana Nikcevic, Maja Stojiljkovic, Branka Zukic, Milena Radmilovic, Teodora Karan-Djurasevic, Sanja Srzentic & Sonja Pavlovic

  2. School of Medicine, University of Belgrade, Belgrade, Serbia

    Milica Colovic

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  1. Vesna Spasovski
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  2. Natasa Tosic
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  3. Gordana Nikcevic
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  4. Maja Stojiljkovic
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  5. Branka Zukic
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  6. Milena Radmilovic
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  7. Teodora Karan-Djurasevic
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  8. Sanja Srzentic
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  9. Milica Colovic
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  10. Sonja Pavlovic
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Corresponding author

Correspondence to Sonja Pavlovic.

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Spasovski, V., Tosic, N., Nikcevic, G. et al. The influence of novel transcriptional regulatory element in intron 14 on the expression of Janus kinase 2 gene in myeloproliferative neoplasms. J Appl Genetics 54, 21–26 (2013). https://doi.org/10.1007/s13353-012-0125-x

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  • DOI: https://doi.org/10.1007/s13353-012-0125-x

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