Person: TURAN, KADİR
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TURAN
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KADİR
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Publication Metadata only The Human MTCH2 Protein Has a Negative Effect On the Influenza a Virus Replication(2021-12-26) TURAN, KADİR; Ulupınar P., Çağlayan E., Turan K.Influenza A viruses have a segmented genome consists of eight single-stranded RNA molecules. Viral replication and transcription are carried out by viral RNA polymerase (RdRP), which consists of PB2, PB2 and PA subunits. During viral infection, RdRP and other viral proteins interact with several host proteins to perform their functions. In this work, mitochondrial carrier homolog 2 (MTCH2) protein, which was found to be associated with viral PA protein by yeast two-hybrid assay, was investigated its importance in terms of viral replication. In order to detect the effects of MTCH2 on virus replication in HeLa and HEK293 cells, the protein level was artificially changed. RNA interference and CRISPR/Cas9 techniques were used for down-regulation of the MTCH2. To increase the MTCH2, the HEK293 cells were transfected with pCHA-MTCH2 plasmid. The cells with altered MTCH2 transcript/protein level were infected with influenza A/WSN and A/DkPen viruses, and the viral replication was evaluated by detection of viral RNA/proteins with qPCR/Western blot techniques and plaque assays. In addition, the RdRP activities in these cells were determined by mini-replicon tests. A significant increase of viral mRNA/protein was observed in knockdown HeLa. It was shown that avian influenza A/DkPen replication was affected more. The over-expression of MTCH2 in HEK293 cells had a negative effect on the viral RdRP. These results showed that the MTCH2 is a negative cellular factor for influenza A virus. Although deletion of MTCH2 genes was detected in HEK293 cells knocked out by CRISPR/Cas9 technique, no negative effect on viral replication was observed. This situation is thought to be caused by heterozygosity or some other factors. From these results, it was concluded that the MTCH2 protein functions as a negative regulation factor for influenza A viruses. This work was supported by a grant of the Scientific and Technological Research Council of Turkey (SBAG-112S518).Keywords: Influenza a Viruses, MTCH2 Protein, Viral Rna Polymerase, PA ProteinPublication Metadata only TÜSEB destekli COVID-19 aşı projeleri(2021-09-09) TURAN, KADİR; Turan K.Publication Open Access Effects of some interferon-related proteins on influenza a viruse RNA polymerase activity(2022-01-01) TURAN, KADİR; Çağlayan E., TURAN K.© 2022, Turkish Pharmacists Association. All rights reserved.Objectives: Interferons (IFNs) are one of the most important components of innate immunity against viruses, especially those carrying the RNA genomes such as influenza viruses. Upon viral infection, the IFNs are rapidly secreted, inducing the expression of several genes in the target cells and establishing an antiviral state. In this study, the effects of proteins encoded by some IFN-related genes on influenza A virus RNA-dependent RNA polymerase enzyme were investigated. We evaluated the importance of these proteins in the pathogenesis of different influenza A virus types. Materials and Methods: The IFN-related genes were amplified by polymerase chain reaction from the HEK293 cDNA library and cloned into pCHA expression vector. The expression of genes and subcellular localizations of the proteins were determined by Western blotting and immunofluorescence staining, respectively. The effects of IFNs-related proteins on virus RdRP enzyme were determined by influenza A virus mini-replicons. Results: The study revealed that the influenza A virus infections significantly altered the transcript level of the IFN-related CCL5, IFIT1, IFIT3, IFITM3, and OAS1 genes in HEK293 cells. It was determined that the alteration of the gene expression was also related to the virus type. The mini-replicon assays showed that the transient expression of CCL5, IFI27, OAS1, IFITM3, IFIT1, and IFIT3 have inhibitory effects on WSN and/or DkPen type virus RdRP enzymes. We observed that the proteins except OAS1 inhibited WSN type RdRP enzyme at a higher level than that of DkPen enzyme. Conclusion: It was concluded that influenza A virus infection significantly alters the IFN-related gene expression in the cells. Most of the proteins encoded from these genes showed an inhibitory effect on the virus RdRP enzymes in the HEK293 cells. The inhibition of the influenza virus RdRP with IFN-related proteins may be the result of direct or indirect interactions between the host proteins and the viral enzyme subunits.Publication Open Access Mutations in the SARS CoV2 spike gene and their reflections on the spike protein(2022-06-01) TURAN, KADİR; Caglayan E., TURAN K.Objective: In this study, it was aimed to determine the mutation frequency in spike (S) genes of SARS CoV2 from six different regions of the world, their distribution on the gene and reflections of these mutations to the S protein. Materials and methods: SARS CoV2 S gene sequences originating from Asia, Africa, Europe, South America, Oceania and North America were obtained from NCBI virus database. The sequences were analyzed with Geneious and BioEdit multiple sequence alignment programs. Results and Conclusion: 865 distinct mutations on the S genes were detected in the virus samples. Among these, 59 variants with numbers of 10 and above in the virus population were detected. The D614G(A1841G) substitution was found to be the most common with an average of 88.6%. Furthermore, it was determined that S477N(G1430A) substitution in theviruses of Oceania differed from other regions with a rate of 86.7%. The average mutation frequency of the S genes from different regions was calculated as 4x10-5. The amino acid substitutions particularly in the RBD (receptor binding domain) have great importance for the virus adsorption to the cells via ACE2 (angiotensin converting enzyme 2) receptor and transmission. However, the importance of these mutations needs to be demonstrated both in silico and experimental studies.Publication Open Access Psychometric properties of the Turkish version of the fear avoidance components scale in patients with chronic pain related to musculoskeletal disorders(2023-01-01) TURAN, KADİR; SARI, ZÜBEYİR; TURAN K., SARI Z., Özden F.Background: The fear avoidance components scale (FACS) evaluates patients’ cognitive, emotional and behavioral fear avoidance behavior. The aim of the study was to conduct the cross-cultural adaptation, reliability and validity of the Turkish version of the FACS. Methods: A prospective cross-sectional study was carried out with 208 patients (46.2 ± 11.4 years, 116 women, 92 men) diagnosed with chronic pain related to musculoskeletal disorders. Individuals were assessed with FACS, Tampa scale of kinesiophobia (TSK), Beck depression inventory (BDI), Oswestry disability index (ODI), numerical pain scale (NPS), and pain catastrophizing scale (PCS). A total of 70 patients completed the FACS for the second time 3 days later. Results: Internal consistency of the total score was excellent (Cronbach’s alpha: 0.815). There was a strong correlation between FACS and TSK and PCS (r1 0.555, r2 0.678, p < 0.001). In addition, the relationship between FACS and BDI and NPS was moderate in terms of construct validity (r1 0.357, r2 0.391, p < 0.001). FACS had a two-factor structure, as expected. The test-retest reliability of the FACS was acceptable to excellent (ICC = 0.526–0.971). Conclusion: The Turkish version of FACS is a valid and reliable questionnaire for patients with chronic pain related to musculoskeletal disorders. The FACS provides a further advantage over identical questionnaires by evaluating cognitive, behavioral and emotional fear avoidance components.