Functional characterization of novel variants in the dnai1 gene in a patient with primary ciliary dyskinesia
Аутори
Skakić, AnitaStevanović, Nina
Spasovski, Vesna
Parezanović, Marina
Ugrin, Milena
Komazec, Jovana
Klaassen, Kristel
Stanković, Sara
Pavlović, Sonja
Stojiljković, Maja
Anđelković, Marina
Конференцијски прилог (Објављена верзија)
Метаподаци
Приказ свих података о документуАпстракт
Primary ciliary dyskinesia (PCD) is a rare motor ciliopathy, which predominantly affects the lungs and
reproductive organs. PCD has a heterogeneous genetic basis, and it is necessary to analyze more than
40 causative genes in order to establish a precise diagnosis, which is essential for optimal treatment and
adequate genetic counseling. Five patients suspected of PCD were analyzed using next-generation
sequencing (NGS). The pathogenicity of the genetic variants was tested by in silico, qRT-PCR and
Western blot methods. Two newly discovered variants p.N450Lfs*6 and p.D562N in the DNAI1 gene
were detected in one patient suspected of PCD. The results of in silico prediction showed that the
p.N450Lfs*6 variant affects the structure of the 3D model of the protein, abolishes ligand binding sites
and post-translational modifications, thereby disrupting protein-protein interactions (PPI). The
p.D562N variant has no effect on the 3D structure of the protein, but affects the ligand bin...ding site and
is located in the WD-40 domain, which most likely disrupts PPI. The results of the qRT-PCR method
showed a decreased expression level of DNAI1 mRNA by about 50% in the patient compared to the
control group, while Western blot analysis showed the presence of two protein products (699 ak and
455 ak). By analyzing the obtained results, it was concluded that the changes p.N450Lfs*6 and
p.D562N affect the length and quantity of the DNAI1 protein, leading to the loss of protein function
and are responsible for the occurrence of primary ciliary dyskinesia in the analyzed patient.
Кључне речи:
PCD / DNAI1 / NGS / qRT-PCR / Western BlotИзвор:
Genetics & Applications, 2023, 7, 2 (Special edition), 110-Издавач:
- Sarajevo : Institute for Genetic Engineering and Biotechnology, University of Sarajevo
Институција/група
Institut za molekularnu genetiku i genetičko inženjerstvoTY - CONF AU - Skakić, Anita AU - Stevanović, Nina AU - Spasovski, Vesna AU - Parezanović, Marina AU - Ugrin, Milena AU - Komazec, Jovana AU - Klaassen, Kristel AU - Stanković, Sara AU - Pavlović, Sonja AU - Stojiljković, Maja AU - Anđelković, Marina PY - 2023 UR - https://imagine.imgge.bg.ac.rs/handle/123456789/1903 AB - Primary ciliary dyskinesia (PCD) is a rare motor ciliopathy, which predominantly affects the lungs and reproductive organs. PCD has a heterogeneous genetic basis, and it is necessary to analyze more than 40 causative genes in order to establish a precise diagnosis, which is essential for optimal treatment and adequate genetic counseling. Five patients suspected of PCD were analyzed using next-generation sequencing (NGS). The pathogenicity of the genetic variants was tested by in silico, qRT-PCR and Western blot methods. Two newly discovered variants p.N450Lfs*6 and p.D562N in the DNAI1 gene were detected in one patient suspected of PCD. The results of in silico prediction showed that the p.N450Lfs*6 variant affects the structure of the 3D model of the protein, abolishes ligand binding sites and post-translational modifications, thereby disrupting protein-protein interactions (PPI). The p.D562N variant has no effect on the 3D structure of the protein, but affects the ligand binding site and is located in the WD-40 domain, which most likely disrupts PPI. The results of the qRT-PCR method showed a decreased expression level of DNAI1 mRNA by about 50% in the patient compared to the control group, while Western blot analysis showed the presence of two protein products (699 ak and 455 ak). By analyzing the obtained results, it was concluded that the changes p.N450Lfs*6 and p.D562N affect the length and quantity of the DNAI1 protein, leading to the loss of protein function and are responsible for the occurrence of primary ciliary dyskinesia in the analyzed patient. PB - Sarajevo : Institute for Genetic Engineering and Biotechnology, University of Sarajevo C3 - Genetics & Applications T1 - Functional characterization of novel variants in the dnai1 gene in a patient with primary ciliary dyskinesia IS - 2 (Special edition) SP - 110 VL - 7 UR - https://hdl.handle.net/21.15107/rcub_imagine_1903 ER -
@conference{ author = "Skakić, Anita and Stevanović, Nina and Spasovski, Vesna and Parezanović, Marina and Ugrin, Milena and Komazec, Jovana and Klaassen, Kristel and Stanković, Sara and Pavlović, Sonja and Stojiljković, Maja and Anđelković, Marina", year = "2023", abstract = "Primary ciliary dyskinesia (PCD) is a rare motor ciliopathy, which predominantly affects the lungs and reproductive organs. PCD has a heterogeneous genetic basis, and it is necessary to analyze more than 40 causative genes in order to establish a precise diagnosis, which is essential for optimal treatment and adequate genetic counseling. Five patients suspected of PCD were analyzed using next-generation sequencing (NGS). The pathogenicity of the genetic variants was tested by in silico, qRT-PCR and Western blot methods. Two newly discovered variants p.N450Lfs*6 and p.D562N in the DNAI1 gene were detected in one patient suspected of PCD. The results of in silico prediction showed that the p.N450Lfs*6 variant affects the structure of the 3D model of the protein, abolishes ligand binding sites and post-translational modifications, thereby disrupting protein-protein interactions (PPI). The p.D562N variant has no effect on the 3D structure of the protein, but affects the ligand binding site and is located in the WD-40 domain, which most likely disrupts PPI. The results of the qRT-PCR method showed a decreased expression level of DNAI1 mRNA by about 50% in the patient compared to the control group, while Western blot analysis showed the presence of two protein products (699 ak and 455 ak). By analyzing the obtained results, it was concluded that the changes p.N450Lfs*6 and p.D562N affect the length and quantity of the DNAI1 protein, leading to the loss of protein function and are responsible for the occurrence of primary ciliary dyskinesia in the analyzed patient.", publisher = "Sarajevo : Institute for Genetic Engineering and Biotechnology, University of Sarajevo", journal = "Genetics & Applications", title = "Functional characterization of novel variants in the dnai1 gene in a patient with primary ciliary dyskinesia", number = "2 (Special edition)", pages = "110", volume = "7", url = "https://hdl.handle.net/21.15107/rcub_imagine_1903" }
Skakić, A., Stevanović, N., Spasovski, V., Parezanović, M., Ugrin, M., Komazec, J., Klaassen, K., Stanković, S., Pavlović, S., Stojiljković, M.,& Anđelković, M.. (2023). Functional characterization of novel variants in the dnai1 gene in a patient with primary ciliary dyskinesia. in Genetics & Applications Sarajevo : Institute for Genetic Engineering and Biotechnology, University of Sarajevo., 7(2 (Special edition)), 110. https://hdl.handle.net/21.15107/rcub_imagine_1903
Skakić A, Stevanović N, Spasovski V, Parezanović M, Ugrin M, Komazec J, Klaassen K, Stanković S, Pavlović S, Stojiljković M, Anđelković M. Functional characterization of novel variants in the dnai1 gene in a patient with primary ciliary dyskinesia. in Genetics & Applications. 2023;7(2 (Special edition)):110. https://hdl.handle.net/21.15107/rcub_imagine_1903 .
Skakić, Anita, Stevanović, Nina, Spasovski, Vesna, Parezanović, Marina, Ugrin, Milena, Komazec, Jovana, Klaassen, Kristel, Stanković, Sara, Pavlović, Sonja, Stojiljković, Maja, Anđelković, Marina, "Functional characterization of novel variants in the dnai1 gene in a patient with primary ciliary dyskinesia" in Genetics & Applications, 7, no. 2 (Special edition) (2023):110, https://hdl.handle.net/21.15107/rcub_imagine_1903 .