The influence of novel transcriptional regulatory element in intron 14 on the expression of Janus kinase 2 gene in myeloproliferative neoplasms
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2013
Authors
Spasovski, VesnaTošić, Nataša
Nikčević, Gordana
Stojiljković, Maja
Zukić, Branka
Ugrin, Milena
Karan-Đurašević, Teodora
Srzentić Dražilov, Sanja
Colović, Milica
Pavlović, Sonja
Article (Published version)
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Show full item recordAbstract
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 gt 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 gt C acts as a transcriptional repressor in vitro. The repressor activity was significantly atte...nuated 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:
Transcriptional regulation / Myeloproliferative neoplasms / JAK2 intron 14 / JAK2 gene expression / 46/1 haplotypeSource:
Journal of Applied Genetics, 2013, 54, 1, 21-26Publisher:
- Springer Heidelberg, Heidelberg
Funding / projects:
- Rare Diseases:Molecular Pathophysiology, Diagnostic and Therapeutic Modalities and Social, Ethical and Legal Aspects (RS-MESTD-Integrated and Interdisciplinary Research (IIR or III)-41004)
DOI: 10.1007/s13353-012-0125-x
ISSN: 1234-1983
PubMed: 23188718
WoS: 000313737500003
Scopus: 2-s2.0-84872673955
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Institut za molekularnu genetiku i genetičko inženjerstvoTY - JOUR AU - Spasovski, Vesna AU - Tošić, Nataša AU - Nikčević, Gordana AU - Stojiljković, Maja AU - Zukić, Branka AU - Ugrin, Milena AU - Karan-Đurašević, Teodora AU - Srzentić Dražilov, Sanja AU - Colović, Milica AU - Pavlović, Sonja PY - 2013 UR - https://imagine.imgge.bg.ac.rs/handle/123456789/619 AB - 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 gt 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 gt 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. PB - Springer Heidelberg, Heidelberg T2 - Journal of Applied Genetics T1 - The influence of novel transcriptional regulatory element in intron 14 on the expression of Janus kinase 2 gene in myeloproliferative neoplasms EP - 26 IS - 1 SP - 21 VL - 54 DO - 10.1007/s13353-012-0125-x ER -
@article{ author = "Spasovski, Vesna and Tošić, Nataša and Nikčević, Gordana and Stojiljković, Maja and Zukić, Branka and Ugrin, Milena and Karan-Đurašević, Teodora and Srzentić Dražilov, Sanja and Colović, Milica and Pavlović, Sonja", year = "2013", 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 gt 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 gt 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.", publisher = "Springer Heidelberg, Heidelberg", journal = "Journal of Applied Genetics", title = "The influence of novel transcriptional regulatory element in intron 14 on the expression of Janus kinase 2 gene in myeloproliferative neoplasms", pages = "26-21", number = "1", volume = "54", doi = "10.1007/s13353-012-0125-x" }
Spasovski, V., Tošić, N., Nikčević, G., Stojiljković, M., Zukić, B., Ugrin, M., Karan-Đurašević, T., Srzentić Dražilov, S., Colović, M.,& Pavlović, S.. (2013). The influence of novel transcriptional regulatory element in intron 14 on the expression of Janus kinase 2 gene in myeloproliferative neoplasms. in Journal of Applied Genetics Springer Heidelberg, Heidelberg., 54(1), 21-26. https://doi.org/10.1007/s13353-012-0125-x
Spasovski V, Tošić N, Nikčević G, Stojiljković M, Zukić B, Ugrin M, Karan-Đurašević T, Srzentić Dražilov S, Colović M, Pavlović S. The influence of novel transcriptional regulatory element in intron 14 on the expression of Janus kinase 2 gene in myeloproliferative neoplasms. in Journal of Applied Genetics. 2013;54(1):21-26. doi:10.1007/s13353-012-0125-x .
Spasovski, Vesna, Tošić, Nataša, Nikčević, Gordana, Stojiljković, Maja, Zukić, Branka, Ugrin, Milena, Karan-Đurašević, Teodora, Srzentić Dražilov, Sanja, Colović, Milica, Pavlović, Sonja, "The influence of novel transcriptional regulatory element in intron 14 on the expression of Janus kinase 2 gene in myeloproliferative neoplasms" in Journal of Applied Genetics, 54, no. 1 (2013):21-26, https://doi.org/10.1007/s13353-012-0125-x . .