• Title/Summary/Keyword: mutational analysis

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A mutational anlaysis of the N-terminal protease of bovine viral diarrhea virus

  • Chon, Seung-ki
    • Korean Journal of Veterinary Research
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    • v.39 no.4
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    • pp.772-777
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    • 1999
  • The uncaped genomic RNA of bovine viral diarrhea virus (BVDV) initiates translation by recruitment of eukaryotic translation initiation factors at the internal ribosome entry site (IRES). N-terminal protease ($N^{pro}$) is the first translation product of the open reading frame (ORF). By using the vaccinia virus SP6 RNA polymerase transient expression system, we showed previously that deletion of $N^{pro}$ region reduced translation by 21%. To better understand the biological significance of $N^{pro}$ for translation, we carried out a mutational analysis of the $N^{pro}$ region of BVDV cloned in the intercistronic region of a bicistronic reporter plasmid. We constructed a bicistronic expression vector in which the entire 5 UTR and the mutated $N^{pro}$ region (${\Delta}386-901$, ${\Delta}415-901$ and ${\Delta}657-901$) was cloned between two reporter genes, chloramphenicol acetyltransferase (CAT) and luciferase (LUC). In vivo translation analyses showed that $N^{pro}$ region was dispensible for efficient translation. The results indicate that the $N^{pro}$ region is not essential for BVDV RNA translation and the 3' boundary of BVDV IRES is expanded into $N^{pro}$ region, suggesting that $N^{pro}$ may not play a major role in BVDV replication.

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Identification of the+1 Ribosomal Frameshifting Site of LRV1-4 by Mutational Analysis

  • Kim Se Na;Choi Jung Ho;Park Min Woo;Jeong Sun Joo;Han Kyung Sook;Kim Hong Jin
    • Archives of Pharmacal Research
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    • v.28 no.8
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    • pp.956-962
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    • 2005
  • Leishmania virus (LRV)1-4 has been reported to produce a fusion of ORF2 and ORF3 via a programmed +1 frameshift in the region where ORF2 and ORF3 overlap (Lee et a/., 1996). However, the exact frameshift site has not been identified. In this study, we compared the frameshift efficiency of a 259bp (nt. 2565-2823), frameshift region of LRV1-4, and the 71 bp (nt. 2605-2678) sub-region where ORF2 and ORF3 overlap. We then predicted the frameshift site using a new computer program (Pseudoviewer), and finally identified the specific region associated with the mechanism of the LRV1-4's+1 frameshift by means of a mutational analysis based on the predicted structure of LRV1-4 RNA. The predicted structure was confirmed by biochemical analysis. In order to measure the frameshift efficiency, constructs that generate luciferase without a frameshift or with a+1 frameshift, were generated and in vitro transcription/translation analysis was performed. Measurements of the luciferase activity generated, showed that the frameshift efficiency was about $1\%$ for both the 259bp (LRV1-4 259FS) and 71 bp region (LRV1-4 71FS). Luciferase activity was strongly reduced in a mutant (LRV1-4 NH: nt. 2635-2670) with the entire hairpin deleted and in a mutant (LRV1-4 NUS: nt. 2644-2659) with the upper stem of the hairpin deleted. These results indicate that the frameshift site in LRV1-4's is in the 71 bp region where ORF2 and ORF3 overlap, and that nt. 2644-2659 (the upward hairpin stem) playa key role in generating the +1 frameshift.

Genetic Features of Lung Adenocarcinoma with Ground-Glass Opacity: What Causes the Invasiveness of Lung Adenocarcinoma?

  • Kim, Dohun;Lee, Jong-Young;Yoo, Jin Young;Cho, Jun Yeun
    • Journal of Chest Surgery
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    • v.53 no.5
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    • pp.250-257
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    • 2020
  • Background: Lung adenocarcinoma (LUAD) with ground-glass opacity (GGO) can become aggravated, but the reasons for this aggravation are not fully understood. The goal of this study was to analyze the genetic features and causes of progression of GGO LUAD. Methods: LUAD tumor samples and normal tissues were analyzed using an Illumina HiSeq 4000 system. After the tumor mutational burden (TMB) was calculated, the identified mutations were classified as those found only in GGO LUAD, those present only in nonGGO LUAD, and those common to both tissue types. Ten high-frequency genes were selected from each domain, after which protein interaction network analysis was conducted. Results: Overall, 227 mutations in GGO LUAD, 212 in non-GGO LUAD, and 48 that were common to both tumor types were found. The TMB was 8.8 in GGO and 7.8 in non-GGO samples. In GGO LUAD, mutations of FCGBP and SFTPA1 were identified. FOXQ1, IRF5, and MAGEC1 mutations were common to both types, and CDC27 and NOTCH4 mutations were identified in the non-GGO LUAD. Protein interaction network analysis indicated that IRF5 (common to both tissue types) and CDC27 (found in the non-GGO LUAD) had significant biological functions related to the cell cycle and proliferation. Conclusion: In conclusion, GGO LUAD exhibited a higher TMB than non-GGO LUAD. No clinically meaningful mutations were found to be specific to GGO LUAD, but mutations involved in the epithelial-mesenchymal transition or cell cycle were found in both tumor types and in non-GGO tissue alone. These findings could explain the non-invasiveness of GGO-type LUAD.

Codon Usage Bias and Determining Forces in Taenia solium Genome

  • Yang, Xing;Ma, Xusheng;Luo, Xuenong;Ling, Houjun;Zhang, Xichen;Cai, Xuepeng
    • Parasites, Hosts and Diseases
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    • v.53 no.6
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    • pp.689-697
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    • 2015
  • The tapeworm Taenia solium is an important human zoonotic parasite that causes great economic loss and also endangers public health. At present, an effective vaccine that will prevent infection and chemotherapy without any side effect remains to be developed. In this study, codon usage patterns in the T. solium genome were examined through 8,484 protein-coding genes. Neutrality analysis showed that T. solium had a narrow GC distribution, and a significant correlation was observed between GC12 and GC3. Examination of an NC (ENC vs GC3s)-plot showed a few genes on or close to the expected curve, but the majority of points with low-ENC (the effective number of codons) values were detected below the expected curve, suggesting that mutational bias plays a major role in shaping codon usage. The Parity Rule 2 plot (PR2) analysis showed that GC and AT were not used proportionally. We also identified 26 optimal codons in the T. solium genome, all of which ended with either a G or C residue. These optimal codons in the T. solium genome are likely consistent with tRNAs that are highly expressed in the cell, suggesting that mutational and translational selection forces are probably driving factors of codon usage bias in the T. solium genome.

A Study on the Correlation between Body-Size and MDVP Parameters in the Normal Male and Female Korean Population (정상 한국인의 성별 체형정보와 MDVP 변수간의 상관관계 연구)

  • Kang, Jae-Hwan;Yoo, Jong-Hyang;Kim, Jong-Yeol
    • Speech Sciences
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    • v.15 no.4
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    • pp.107-119
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    • 2008
  • This paper intends to investigate the correlation of 12 MDVP measurements with age, sex and body-size of sampled healthy patients. In order to extract pitch and 12 MDVP parameters efficiently and display the correlation of each parameter easily, we developed the speech analysis program using C/C++ and MFC development tool. The sample group consists of 205 males and 343 females with ages 9-81. We collected vowel voices /a/ and 8 body-size measurements from them. Body-size values were taken at 8 different torso positions of each person. We analyzed the matched voice samples and body-size measurements by the developed speech analysis program and SPSS program. The result shows that a typical characteristic age-F0 pattern that F0 of male subjects are rapidly decreased after mutational period and have stable state with age and that of female subjects are slowly changed by overall age. In MDVP, age-STD in males, age-sPPQ in females relationships are especially similar to the age-F0 relationship. In case of male group, sPPQ(0.316%), Jitt(0.04%), Shim(0.25%), APQ(0.28%) variables are increased with age after mutational period. And Jitt(0.042%), sPPQ(0.219%) of females group are increased with age too. In cases of height, weight and BMI there exists a weak correlation with MDVP, which shows a correlation coefficient below 0.25 about male and female groups. The survey of correlation relationship between 8 body-size measurements and MDVP shows a insignificant statistical result by only just having the correlation coefficient maximum in M8-8 and F0(-0.394%) for males and M8-6,7(-0.368%, -0.364%) for females.

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Annotation of Genes Having Candidate Somatic Mutations in Acute Myeloid Leukemia with Whole-Exome Sequencing Using Concept Lattice Analysis

  • Lee, Kye Hwa;Lim, Jae Hyeun;Kim, Ju Han
    • Genomics & Informatics
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    • v.11 no.1
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    • pp.38-45
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    • 2013
  • In cancer genome studies, the annotation of newly detected oncogene/tumor suppressor gene candidates is a challenging process. We propose using concept lattice analysis for the annotation and interpretation of genes having candidate somatic mutations in whole-exome sequencing in acute myeloid leukemia (AML). We selected 45 highly mutated genes with whole-exome sequencing in 10 normal matched samples of the AML-M2 subtype. To evaluate these genes, we performed concept lattice analysis and annotated these genes with existing knowledge databases.

Effects of a Phosphomimetic Mutant of RAP80 on Linear Polyubiquitin Binding Probed by Calorimetric Analysis

  • Thach, Thanh Trung;Jee, Jun-Goo;Lee, Sang-Ho
    • Bulletin of the Korean Chemical Society
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    • v.33 no.4
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    • pp.1285-1289
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    • 2012
  • RAP80 plays a key role in DNA damage responses by recognizing K63-linked polyubiquitin moieties through its two ubiquitin-interacting motif (UIM) domains. The linker between the two UIMs possesses a phosphorylation site, but the relationship between phosphorylation and polyubiquitin recognition remains elusive. We investigated the interaction between a phosphorylation-mimic RAP80 mutant S101E and linear polyubiquitins, structurally equivalent to the K63-linked ones, using isothermal titration calorimetry (ITC). ITC analysis revealed differential binding affinities for linear tetraubiquitin by otherwise equivalent UIMs in S101E. Mutational analysis supported such differential polyubiquitin recognition by S101E. Our results suggest a potential crosstalk between polyubiquitin recognition and phosphorylation in RAP80.

Acquired JAK-2 V617F Mutational Analysis in Pakistani Patients with Essential Thrombocythemia

  • Sultan, Sadia;Irfan, Syed Mohammed
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.16
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    • pp.7327-7330
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    • 2015
  • Background: Essential thrombocythemia (ET) is a clonal hemopoietic stem cell myeloproliferative neoplasm characterized by persistent thrombocytosis along with megakaryocytic hyperplasia. In the last decade following the identification of an acquired JAK2 V617F mutation, there has been acceleration in our understanding of this disease. The rational of this study was to determine the mutational profile of JAK2 V617F in Pakistan patients with ET. Materials and Methods: In this retrospective cross sectional study, 21 patients with ET were enrolled from January 2011 to December 2014. Patients were diagnosed based on WHO criteria for essential thrombocythemia. Complete blood count was done on an automated hematology analyzer, while JAK2 V617F expression was evaluated by polymerase chain reaction. Results: The mean age was $56.7{\pm}19.0$ years (range 18-87) and the male to female ratio was 1:1.1. The frequency of JAK2 V617F positivity in our ET patients was found to be 61.9%. The mean hemoglobin was $11.7{\pm}2.4$ g/dl with a total leukocyte count of $13.3{\pm}8.1{\times}109/l$ and a platelet count of $1188{\pm}522{\times}109/l$. Positive correlations for JAK2 V617F mutation were established with high TLC count and raised LDH (P<0.05). No correlation of JAK2 V617F could be established with age and gender (P>0.05). Conclusions: JAK2 V617F mutation frequency in our ET patients was similar to those reported previously. Screening for the mutation in all suspected essential thrombocythemia cases could be beneficial in differentiating patients with reactive and clonal thrombocytosis.

Somatic mutation patterns and compound response in cancers

  • He, Ningning;Kim, Nayoung;Yoon, Sukjoon
    • BMB Reports
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    • v.46 no.2
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    • pp.97-102
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    • 2013
  • The use of various cancer cell lines can recapitulate known tumor-associated mutations and genetically define cancer subsets. This approach also enables comparative surveys of associations between cancer mutations and drug responses. Here, we analyzed the effects of ~40,000 compounds on cancer cell lines that showed diverse mutation-dependent sensitivity profiles. Over 1,000 compounds exhibited unique sensitivity on cell lines with specific mutational genotypes, and these compounds were clustered into six different classes of mutation-oriented sensitivity. The present analysis provides new insights into the relationship between somatic mutations and selectivity response of chemicals, and these results should have applications related to predicting and optimizing thera-peutic windows for anti-cancer agents.