Journal of Applied Genetics

, Volume 54, Issue 1, pp 21–26

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

  • Vesna Spasovski
  • Natasa Tosic
  • Gordana Nikcevic
  • Maja Stojiljkovic
  • Branka Zukic
  • Milena Radmilovic
  • Teodora Karan-Djurasevic
  • Sanja Srzentic
  • Milica Colovic
  • Sonja Pavlovic
Human Genetics • Original Paper

DOI: 10.1007/s13353-012-0125-x

Cite this article as:
Spasovski, V., Tosic, N., Nikcevic, G. et al. J Appl Genetics (2013) 54: 21. doi:10.1007/s13353-012-0125-x

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.

Keywords

Myeloproliferative neoplasmsJAK2 intron 1446/1 haplotypeJAK2 gene expressionTranscriptional regulation

Copyright information

© Institute of Plant Genetics, Polish Academy of Sciences, Poznan 2012

Authors and Affiliations

  • Vesna Spasovski
    • 1
  • Natasa Tosic
    • 1
  • Gordana Nikcevic
    • 1
  • Maja Stojiljkovic
    • 1
  • Branka Zukic
    • 1
  • Milena Radmilovic
    • 1
  • Teodora Karan-Djurasevic
    • 1
  • Sanja Srzentic
    • 1
  • Milica Colovic
    • 2
  • Sonja Pavlovic
    • 1
  1. 1.Institute of Molecular Genetics and Genetic EngineeringUniversity of BelgradeBelgradeSerbia
  2. 2.School of MedicineUniversity of BelgradeBelgradeSerbia