• Title/Summary/Keyword: Triplex

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RNA helicase DEAD-box-5 is involved in R-loop dynamics of preimplantation embryos

  • Hyeonji Lee;Dong Wook Han;Seonho Yoo;Ohbeom Kwon;Hyeonwoo La;Chanhyeok Park;Heeji Lee;Kiye Kang;Sang Jun Uhm;Hyuk Song;Jeong Tae Do;Youngsok Choi;Kwonho Hong
    • Animal Bioscience
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    • v.37 no.6
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    • pp.1021-1030
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    • 2024
  • Objective: R-loops are DNA:RNA triplex hybrids, and their metabolism is tightly regulated by transcriptional regulation, DNA damage response, and chromatin structure dynamics. R-loop homeostasis is dynamically regulated and closely associated with gene transcription in mouse zygotes. However, the factors responsible for regulating these dynamic changes in the R-loops of fertilized mouse eggs have not yet been investigated. This study examined the functions of candidate factors that interact with R-loops during zygotic gene activation. Methods: In this study, we used publicly available next-generation sequencing datasets, including low-input ribosome profiling analysis and polymerase II chromatin immunoprecipitation-sequencing (ChIP-seq), to identify potential regulators of R-loop dynamics in zygotes. These datasets were downloaded, reanalyzed, and compared with mass spectrometry data to identify candidate factors involved in regulating R-loop dynamics. To validate the functions of these candidate factors, we treated mouse zygotes with chemical inhibitors using in vitro fertilization. Immunofluorescence with an anti-R-loop antibody was then performed to quantify changes in R-loop metabolism. Results: We identified DEAD-box-5 (DDX5) and histone deacetylase-2 (HDAC2) as candidates that potentially regulate R-loop metabolism in oocytes, zygotes and two-cell embryos based on change of their gene translation. Our analysis revealed that the DDX5 inhibition of activity led to decreased R-loop accumulation in pronuclei, indicating its involvement in regulating R-loop dynamics. However, the inhibition of histone deacetylase-2 activity did not significantly affect R-loop levels in pronuclei. Conclusion: These findings suggest that dynamic changes in R-loops during mouse zygote development are likely regulated by RNA helicases, particularly DDX5, in conjunction with transcriptional processes. Our study provides compelling evidence for the involvement of these factors in regulating R-loop dynamics during early embryonic development.

Symptoms of Cucumber Virus Diseases Occurred in Sangju and Gurye in 2006 and 2007 (2006년과 2007년 상주와 구례에서 발생한 오이 바이러스병의 병징 특성)

  • Cho, Jeom-Deog;Lee, Jung-Hwan;Ko, Sug-Ju;Choi, Hong-Soo;Lee, Su-Heon;Choi, Gug-Seoun;Kim, Jeong-Soo
    • Research in Plant Disease
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    • v.17 no.2
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    • pp.196-204
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    • 2011
  • Virus diseases occurring on cucumber was surveyed at main plantation areas of 'Sangju' and 'Gurye' in 2006 and 2007. Viral infection rate on cucumber was ranged from 14% to 90% in fields and the average infection rate was 46% at Sangju area. Cucumber cultivated at Gurye area had viral incidence ranged 9-100% and averaged 48%. The majorly infected viruses were Cucumber green mottle mosaic virus, Zucchini yellow mosaic virus, Papaya ring spot virus and Watermelon mosaic virus-2, and their infection rates were 23.5%, 13.0%, 9.0% and 2.0%, respectively in 2006 and 2007. Mixed infection rate of duplex, triplex and tetraplex was 31.5%, 7.5% and 2.5%, respectively. The rate of infection type was 47.7% by single infection, 31.5% by double infection and 88.7% by both infection types. Important viruses involved in mixed infection types were Zucchini yellow mosaic virus, Cucumber green mottle mosaic virus, Papaya ring spot virus and Broad been wilt virus2, orderly. Symptom in single infection was almost same on 'Dadagi' line and 'Chicheong' line of cucumber cultivars. Cucumber green mottle mosaic virus produced various symptoms of chlorotic spot, vein chlorosis and vein wrinkle and so on. Main symptoms of vein chlorosis, and severe mosaic and malformation were induced by Zucchini yellow mosaic virus. Mild symptoms were occurred relatively by Papaya ring spot virus and Watermelon mosaic virus2.

Clinical and Phylogenetic Characteristics of Escherichia coli Urinary Tract Infections (대장균에 의한 요로감염의 연령별, 계통군별 특징)

  • Lee, Ji Eun;Lee, Youn Hee;Nam, Chan Hee;Kwak, Ga Young;Lee, Soo Young;Kim, Jong Hyun;Hur, Jae Kyun;Kang, Jin Han
    • Pediatric Infection and Vaccine
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    • v.17 no.1
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    • pp.16-22
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    • 2010
  • Purpose : We aimed to investigate the clinical and phylogenetic characteristics of Escherichia coli Urinary Tract Infections (E. coli UTI). Methods : We enrolled patients with culture-proven E. coli UTI, who were admitted at the study hospital from September 2008 to August 2009. We investigated clinical data of patients with E. coli UTI and characteristics of isolated E. coli strains. The phylogenetic groups were classified using triplex polymerase chain reaction (PCR), and the distribution of nine virulent genes was determined by multiplex PCR. Results : A total of 47 patients have participated in this study. Thirty (63.8%) were under 6 months; eight (17.0%) were between 6-12 months; and nine (19.1%) were over 12 months. We compared two age groups between under 6-month and over 6-month. In the age group under 6-month, higher proportion of male (P =0.002) and group B2 strains (P =0.020) were observed. In contrast, higher proportion of female and group non-B2 strains were observed in age group over 6-month. Frequencies of papC, papGII, papGIII, sfa/foc, hlyC, cnf1, fyuA, iroN and iucC were estimated as 68.1%, 57.4%, 42.6%, 46.8%, 46.8%, 31.9%, 87.2%, 48.9% and 63.8%, respectively. In the comparison of phylogenetic groups, group B2 showed higher distribution of virulent genes, while group D included more strains resistant to trimethoprim/sulfamethoxazole (TMP/SMZ) than other groups. Conclusion : We showed the age group-specific difference in the distribution of sex ratios and phylogenetic groups; more male and group B2 strains in age group under 6-month, while more female and group non-B2 in age group over 6-month. However, further evaluation including larger number of patients will be necessary to confirm above thesis in future molecular epidemiological studies.