• Animal Bioscience
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• 제37권6호
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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.

• 설비공학논문집
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• 제13권8호
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• pp.692-700
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• 2001

Flow boiling heat transfer coefficients(HTCs) of R22, R134a, R407C, and R410A were measured experimentally for a horizontal plain and a microfin tube. Experimental apparatus was composed of 3 main parts: a refrigerant loop, a water loop and a water-glycol loop. The test section in th refrigerant loop was made of a copper tube of 9.52 mm outer diameter and 1 m length for both tubes. The refrigerant was heated by passing hot water through an annulus surrounding the test section. Tests were performed at a fixed refrigerant saturation temperature of $5^{\circ}C$ with mass fluxes of 100~300 kg/$m^2$s. Test results showed that at similar mass flux the flow boiling HTCs of R134a were similar to those of R22 for both plain and microfin tube. HTCs of R407C were similar to those of R22 for a plain tube but lower than those of R2 by 25~48% for a microfin tube. And HTCs of R410A were higher than those of R2 by 20~63% for a plain tube and were similar to those of R22 for a microfin tube. In general, HTCs of a microfin tube were 1.8~5.7 times higher than those of a plain tube.

• 설비공학논문집
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• 제14권2호
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• pp.175-183
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• 2002

Flow Condensation heat transfer coefficients (HTCs) of Rl2, R22, R32, Rl23, Rl25, R134a, R142b were measured experimentally on a horizontal plain tube. The experi- mental apparatus was composed of three main parts; a refrigerant loop, a water loop and a water-glycol loop. The test section in a refrigerant loop was made of a copper tube of 8.8 mm inner diameter and 1000 mm length respectively. The refrigerant was cooled by passing cold water through an annulus surrounding the test section. All tests were performed at a filed refrigerant saturation temperature of 4$0^{\circ}C$ with mass fluxes of 100, 200, 300 kg/$m^2$s. The experimental result showed that flow condensation HTCs increase as the quality, mass flux, and latent heat of condensation increase. At the same mass flux, the HTCs of R32 and R142b were higher than those of R22 by 35~45% and 7~14% respectively while HTCs of R134a and Rl23 were similar to those of R22. On the other hand, HTCs of Rl25 and Rl2 were lower than those of R22 by 28 ~30% and 15 ~25% respectively Finally, a new correlation for flow condensation HTCs was developed by modifying Dobson and Chato's correlation with the latent heat of condensation considered. The correlaton showed an average deviation of 13.1% for all pure fluids data indicating an excellent agreement.

• 대한기계학회:학술대회논문집
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• 대한기계학회 2004년도 추계학술대회
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• pp.1270-1275
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• 2004

Flow condensation heat transfer coefficients(HTCs) of R22 and R134a were measured on horizontal aluminum multi-channel tube. The experimental apparatus was composed of three main parts ; a refrigerant loop, a water loop and a water-ethylene glycol loop. The test section in the refrigerant loop was made of aluminum multi-channel tube of 1.4 mm hydraulic diameter and 0.53 m length. The refrigerant was cooled by passing cold water through an annulus surrounding the test section. The data scan vapor qualities $(0.1{\sim}0.9)$, mass flux ($200{\sim}400$ $kg/m^{2}s$) and heat flux ($7.3{\sim}7.7$ $kW/m^{2}$) at $40{\times}0.2^{\circ}C$ saturation temperature in small hydraulic diameter tube. It was found that some well-known previous correlations were not suitable for multichannel tube. So, It must develop new correlations for multi-channel tubes.

• 설비공학논문집
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• 제16권5호
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• pp.444-451
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• 2004

Flow condensation heat transfer coefficients (HTCs) of R22, R134a, R407C, and R410A were measured on horizontal plain and microfin tubes. The experimental apparatus was composed of three main parts; a refrigerant loop, a water loop and a water/glycol loop. The test section in the refrigerant loop was made of both a plain and a microfin copper tube of 6.0∼6.16 mm inside diameter and 1.0 m length. Refrigerants were cooled by passing cold water through an annulus surrounding the test section. Tests were performed at a fixed refrigerant saturation temperature of 4$0^{\circ}C$ with mass fluxes of 100, 200, and 300 kg/m2s. Test results showed that at similar mass flux the flow condensation HTCs of R134a were similar to those of R22 for both plain and microfin tubes. On the other hand, HTCs of R407C were lower than those of R22 by 4∼16% and 16∼42% for plain and microfin tubes respectively. And HTCs of R410A were similar to those of R22 for a plain tube but lower than those of R22 by 3∼9% for a microfin tube. Heat transfer enhancement factors of a microfin tube were 1.3∼1.9.

• 생명과학회지
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• 제23권10호
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• pp.1260-1266
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• 2013

본 연구에서는 H. marmoreus 3-10균주와 H. marmoreus 1-1균주의 ribosomal DNA (rDNA) cluster의 분석이 수행되었다. Small subunit (SSU)와 intergenic spacer 2 (IGS 2)는 부분적으로 염기서열이 결정되었고, internal transcribed spacer 1 (ITS 1), 5.8S, internal transcribed spacer 2 (ITS 2), large subunit (LSU), intergenic spacer 1 (IGS 1), 5S는 완전하게 염기서열이 결정 되었다. 팽이버섯 H. marmoreus 3-10균주와 H. marmoreus 1-1균주의 rDNA cluster는 총 7,049 bp로 결정되었다. SSU은 1,796 bp, ITS1은 229 bp, 5.8S은 153 bp, ITS2는 223 bp, LSU은 3,348 bp, IGS1은 390 bp, IGS2은 900 bp로 염기서열이 분석되었다. 결정된 rDNA cluster의 총 7,049 bp 중에서 17 bp가 다름이 확인되었고, 각각 SSU (2 bp), ITS (3 bp), LSU (9 bp), IGS (3 bp)에서 차이를 확인하였다. ITS regions의 2차 구조 결과 5개의 stem-loop가 있음이 드러났다. 흥미롭게도, 이들 stem-loop 사이에서 stem-loop V에서 한 개의 상이한 염기가 다른 2차 구조를 나타냄을 확인하였다.

• 설비공학논문집
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• 제14권8호
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• pp.656-663
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• 2002

Flow condensation heat transfer coefficients (HTCs) of R22, R134a, R407C, and R410A were measured on horizontal plain and microfin tubes. The experimental apparatus was composed of three main parts; a refrigerant loop, a water loop and a water/glycol loop. The test section in the refrigerant loop was made of both a plain and a microfin copper tube of 9.52 mm outside diameter and 1.0 m length. The refrigerant was cooled by passing cold water through an annulus surrounding the test section. Tests were performed at a fixed refrigerant saturation temperature of $40^{\circ}C$ with mass fluxes of 100, 200, and 300 kg/$m^2s$. Test results showed that at similar mass flux the flow condensation HTCs of R134a were similar to those of R22 for both plain and microfin tubes. On the other hand, HTCs of R407C were lower than those of R22 by 11~l5% and 23~53% for plain and microfin tubes respectively. And HTCs of R410A were similar to those of R22 for a plain tube but lower than those of R22 by 10~21% for a microfin tube. In general, HTCs of a microfin tube were 2.0~3.0 times higher than those of a plain tube.

• 대한화학회지
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• 제38권10호
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• pp.769-773
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• 1994

우유에서 추출한 안지오제닌을 사용하여 Xanthomonas celebensis 5S rRNA의 고차원 구조를 조사하였다. 안지오제닌은 5S rRNA의 단일가닥 부분에 있는 피리미딘 염기들의 먼 쪽으로 있는 3' P-O 에스테르 결합들만을 절단하였다. pH 7.0이고 10 mM의 $Mg^{2+}$이 있을 때 안지오제닌은 5S rRNA의 d 고리에서만 작용하였지만 $Mg^{2+}$이 없을 때는 e 고리를 제외한 모든 고리들(a, b, c, d 고리들)에서 작용하였다. $Mg^{2+}$이 없을 때 $U_{74}$-$G_{75}$와 $U_{77}$-$A_{78}$, $U_{103}$-$A_{104}$ 결합들이 pH 7.0과 pH 3.5에서 모두 안지오제닌의 작용에 매우 민감하였다. 한편 pH 3.5이고 $Mg^{2+}$이 없을 때 안지오제닌이 a 고리의 $C_{17}$-$G_{18}$과 b고리의 $U_{55}$-$G_{56}$ 결합들을 강하게 절단하였다. 이러한 결과들에서 우리는 다음과 같이 결론을 내릴 수 있다. 첫째, 안지오제닌을 5S rRNA의 삼차구조 분석에서의 탐침의 하나로 사용할 수 있음을 알았다. 둘째, 5S rRNA의 d고리의 구조는 $Mg^{2+}$과 $H^+$농도에 따라 변하기 쉽다는 것을 알았다.

• 대한화학회지
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• 제37권2호
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• pp.237-243
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• 1993

Xanthomonas celebensis 5S rRNA를 분리 정제하여 효소적 방법과 화학적 방법으로 그 일차구조 및 이차구조를 결정하였다. 이 5S rRNA는 119개의 누클레오티드로 구성되어 있으며 변형된 누클레오시드를 함유하고 있지 않다. 그리고 이 5S rRNA는 X. maltophilia 및 X. citri의 것처럼 5'-말단에 가외의 우리딘잔기를 하나 더 가지고 있다. 결정한 X. celebensis 5S rRNA의 이차구조는 본 연구진이 이미 결정한 두가지 Xanthomonas 종들의 것들과 매우 유사하며, 5개의 이중나선 줄기와 5개의 단일가닥 고리 그리고 2개의 내밀린 구조를 가지고 있다. X.celebensis 5S rRNA 분자 내의 삼차상호작용은 테옥시헥사머를 이용한 혼성체화법과 Fe(II)-EDTA를 사용하여 5S rRNA를 쪼개는 방법으로 분석하였다. 5S rRNA 상의 $U_{35}CCCAU_{40}$부분을 상보성을 가진 데옥시헥사머를 혼성체화한 다음에 고리 M, 구역 $I_1-C$, 고리 $H_2$에 있는 약간의 아데노신 잔기들이 피로탄산 이에틸에 대한 감수성을 가지게 되는 사실과, $Mg^{2+}$이 있는 조건에서 고리 M, 고리 $H_1$, 및 구역 $D-I_2에$ 있는 약간의 누클레오티드 잔기들이 Fe(II)-EDTA의 분열작용에 저항성을 나타내는 사실에서 고리 $H_1과$ 고리 $H_2$가 어떤 방법으로든 고리 M과 삼차상호작용을 하는 것으로 추측되며, 이 삼차작용에서 구역 $I_1-C$와 구역 $D-I_2$ 돌쩌귀로 작용하는 것으로 생각된다. 고리 $H_1$은 산성 조건(pH 5.5)에서 비표준형 염기쌍 A:C 들을 형성함으로써 3개의 염기로 이루어지는 고리를 형성한다는 것을 알았다.

• 한국생물물리학회:학술대회논문집
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• 한국생물물리학회 2003년도 정기총회 및 학술발표회
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• pp.66-66
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• 2003

In skeletal muscle cells, depolarization of the transverse tubules (T-tubules) results in Ca$\^$2+/ release from the sarcoplasmic reticulum (SR), leading to elevated cytoplasmic Ca$\^$2+/ and muscle contraction. This process has been known as excitation-contraction coupling (E-C coupling). Several proteins, such as the ryanodine receptor (RyR), triadin, junctin, and calsequestrin (CSQ), have been identified to be involved in the Ca$\^$2＋/ release process. However, the molecular interactions between the SR proteins have not been resolved. In the present study, the mechanisms of interaction between RyRl and triadin have been studied by in vitro protein binding and $\^$45/Ca$\^$2+/ overlay assays. Our data demonstrate that the intraluminal loop II of RyR1 binds to triadin in Ca$\^$2+/-independent manner. Moreover, we could not find any Ca$\^$2+/ binding sites in the loop II region. GST-pull down assay revealed that a KEKE motif of triadin, which was previously identified as a CSQ binding site (Kobayasi et al.,2000 JBC) was also a binding site for RyR1. Our results suggest that the intraluminal loop II of RyR could participate in the RyR-mediated Ca$\^$2+/ release process by offering a direct binding site to luminal triadin.