• Reproductive and Developmental Biology
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• 제38권2호
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• pp.63-70
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• 2014

Plasminogen activators (PAs) are serine proteases that convert plasminogen to plasmin. Two type of PAs are urokinase-type PA (uPA) and tissue-type PA (tPA). Plasminogen is present in most extracellular fluids. PAs play in various reproductive processes including implantation, ovulation and fertilization. In the spermatozoa, PAs and PAIs play a role in sperm motility and fertilization. PAs in the sertoli cell are stimulated spermatozoa maturation and sperm activation through the phospholipase A2. The oocyte maturation is the process for fertilization and implantation. PAs in cumulus-oocyte complexes (COCs) are related to oocyte maturation by protein kinase A and C. In the ovulatory process, PAs activity are changed and it are related to reducing the tensile strength of ovarian follicle wall. The uterine environment is important for reproduction and the uterus undergo tissue remodeling. In the uterus and oviduct of mammals, expression and activity of PAs are changed during estrous cycle. Thus, expression and activity of PAs are concerned to many reproductive functions. Therefore, PAs seem to important factor of regulator in reproductive events.

• 한국응용곤충학회지
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• 제45권3호
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• pp.241-259
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• 2006

폴리드나바이러스는 고치벌 및 맵시벌류에 공생하는 DNA 바이러스로 기주 염색체에 프로바이러스 형태로 존재한다. 이 바이러스의 복제는 기주 용발육시기에 난소받침 상피세포에서 시작되어 유리 바이러스 형태의 입자 구조를 이루게 된다. 바이러스 입자는 기주가 피기생체에 산란할 때 알과 함께 혈강으로 옮겨진다. 이 바이러스 게놈의 염기서열을 바탕으로 여러 폴리드나바이러스 유전자군이 동정되었으며, 이들의 생리적 기능도 알려지고 있다. 본 종설은 기생 생리적 견지에서 폴리드나바이러스 게놈을 특성화하고, 이를 토대로 생리 교란 유전자들을 응용할 수 있는 새로운 해충 방제 전략을 소개한다.

• BMB Reports
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• 제46권2호
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• pp.92-96
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• 2013

The breast cancer susceptibility gene BRCA1 encodes a nuclear protein, which functions as a tumor suppressor and is involved in gene transcription and DNA repair processes. Many families with inherited breast and ovarian cancers have mutations in the BRCA1 gene. However, only a few studies have reported on the mechanism underlying the regulation of BRCA1 expression in humans. In this study, we investigated the transcriptional regulation of BRCA1 in HeLa cells treated with etoposide. We found that three Egr-1-binding sequences (EBSs) were located at -1031, -1005, and -385 within the enhancer region of the BRCA1 gene. Forced expression of Egr-1 stimulated the BRCA1 promoter activity. EMSA data showed that Egr-1 bound directly to the EBS within the BRCA1 gene. Knockdown of Egr-1 through the expression of a small hairpin RNA (shRNA) attenuated etoposide-induced BRCA1 promoter activity. We conclude that Egr-1 targets the BRCA1 gene in HeLa cells exposed to etoposide.

• Molecules and Cells
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• 제28권1호
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• pp.31-36
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• 2009

Centrobin/Nip2 was initially identified as a centrosome protein that is critical for centrosome duplication and spindle assembly. In the present study, we determined the expression and subcellular localization of centrobin in selected mouse tissues. Immunoblot analysis revealed that the centrobin-specific band of 100 kDa was detected in all tissues tested but most abundantly in the thymus, spleen and testis. In the testis, centrobin was localized at the centrosomes of spermatocytes and early round spermatids, but no specific signal was detected in late round spermatids and elongated spermatids. Our results also revealed that the centrosome duplication occurs at interphase of the second meiotic division of the mouse male germ cells. The centrobin protein was more abundant in the mitotically active ovarian follicular cells and thymic cortex cells than in non-proliferating corpus luteal cells and thymic medullary cells. The expression pattern of centrobin suggests that the biological functions of centrobin are related to cell proliferation. Consistent with the proposal, we observed reduction of the centrobin levels when NIH3T3 became quiescent in the serum-starved culture conditions. However, a residual amount of centrobin was also detected at the centrosomes of the resting cells, suggesting its role for maintaining integrity of the centrosome, especially of the daughter centriole in the cells.

• Journal of Microbiology and Biotechnology
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• 제32권5호
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• pp.657-662
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• 2022

Glycosyltransferase (GT)-specific degenerate PCR screening followed by in silico sequence analyses of the target clone was used to isolate a member of family1 GT-encoding genes from the established fosmid libraries of soil actinomycetes Micromonospora echinospora ATCC 27932. A recombinant MeUGT1 was heterologously expressed as a His-tagged protein in E. coli, and its enzymatic reaction with semi-synthetic phenoxodiol isoflavene (as a glycosyl acceptor) and uridine diphosphate-glucose (as a glycosyl donor) created two different glycol-attached products, thus revealing that MeUGT1 functions as an isoflavonoid glycosyltransferase with regional flexibility. Chromatographic separation of product glycosides followed by the instrumental analyses, clearly confirmed these previously unprecedented glycosides as phenoxodiol-4'-α-O-glucoside and phenoxodiol-7-α-O-glucoside, respectively. The antioxidant activities of the above glycosides are almost the same as that of parental phenoxodiol, whereas their anti-proliferative activities are all superior to that of cisplatin (the most common platinum chemotherapy drug) against two human carcinoma cells, ovarian SKOV-3 and prostate DU-145. In addition, they are more water-soluble than their parental aglycone, as well as remaining intractable to the simulated in vitro digestion test, hence demonstrating the pharmacological potential for the enhanced bio-accessibility of phenoxodiol glycosides. This is the first report on the microbial enzymatic biosynthesis of phenoxodiol glucosides.

• Biomolecules & Therapeutics
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• 제31권3호
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• pp.330-339
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• 2023

Liver kinase B1 (LKB1) is a crucial tumor suppressor involved in various cellular processes, including embryonic development, tumor initiation and progression, cell adhesion, apoptosis, and metabolism. However, the precise mechanisms underlying its functions remain elusive. In this study, we demonstrate that LKB1 interacts directly with malic enzyme 3 (ME3) through the N-terminus of the enzyme and identified the binding regions necessary for this interaction. The binding activity was confirmed to promote the expression of ME3 in an LKB1-dependent manner and was also shown to induce apoptosis activity. Furthermore, LKB1 and ME3 overexpression upregulated the expression of tumour suppressor proteins (p53 and p21) and downregulated the expression of antiapoptotic proteins (nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and B-cell lymphoma 2 (Bcl-2)). Additionally, LKB1 and ME3 enhanced the transcription of p21 and p53 and inhibited the transcription of NF-κB. Moreover, LKB1 and ME3 suppressed the phosphorylation of various components of the phosphatidylinositol-4,5-bisphosphate 3-kinase/protein kinase B signaling pathway. Overall, these results suggest that LKB1 promotes pro-apoptotic activities by inducing ME3 expression.

• 한국임상수의학회지
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• 제31권5호
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• pp.394-402
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• 2014

본 연구의 목적은 예측 불가능한 만성적인 스트레스가 생식기능과 황체형성호르몬 수용체의 발현에 미치는 영향을 알아보는 것이다. 9 주령 암컷 C57BL/6 마우스를 무작위로 선택하여 대조군과 스트레스군의 두 집단으로 분류하였다. 스트레스군은 35일 동안 하루에 두 번씩 12가지의 서로 다른 스트레스를 무작위로 선택하여 스트레스를 주었다. 대조군에 비하여 스트레스군에서 불안과 관련된 행동들이 유의성 있게 증가하였으며(P < 0.05), 스트레스를 받는 동안의 증체율 또한 유의성 있게 감소하였다(P < 0.01). 그리고 평균 산자수도 대조군에 비하여 스트레스 군에서 유의하게 감소함을 관찰 하였다(P < 0.01). 조직학적인 검사에서 일차, 이차 및 초기 성숙 난포의 비율이 대조군에 비해 스트레스 군에서 유의하게 감소한 반면(P < 0.05) 폐쇄 난포의 비율은 유의하게 증가하였다(P < 0.01). 면역조직화학적 검사를 통해 과립막세포와 황체세포의 황체형성호르몬 수용체 발현을 관찰한 결과, 대조군에 비해 스트레스군에서 그 발현이 감소하였고, 웨스턴 블롯을 통해 난소 내 황체형성호르몬 수용체의 단백질 양을 측정한 결과 또한 대조군에 비하여 스트레스군에서 유의하게 감소하였다(P < 0.05). 본 연구를 통해 난소의 황체형성호르몬 수용체는 예측 불가능한 스트레스에 의해 영향을 받으며, 변화된 황체형성호르몬 수용체가 난자의 난포 발육 불량과 생식기능의 이상에 영향을 미친다는 것을 실험적으로 증명하였다.

• Asian-Australasian Journal of Animal Sciences
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• 제31권5호
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• pp.650-657
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• 2018

Objective: The study investigated the biological functions and mechanisms for controlling cashmere growth of Liaoning cashmere goat by ovarian carcinoma immunoreactive antigen-like protein 2 (OCIAD2) and decorin (DCN) genes. Methods: cDNA library of Liaoning cashmere goat was constructed in early stages. OCIAD2 and DCN genes related to cashmere growth were identified by homology analysis comparison. The expression location of OCIAD2 and DCN genes in primary and secondary hair follicles (SF) was performed using in situ hybridization. The expression of OCIAD2 and DCN genes in primary and SF was performed using real-time polymerase chain reaction (PCR). Results: In situ hybridization revealed that OCIAD2 and DCN were expressed in the inner root sheath of Liaoning cashmere goat hair follicles. Real-time quantitative PCR showed that these genes were highly expressed in SF during anagen, while these genes were highly expressed in primary hair follicle in catagen phase. Melatonin (MT) inhibited the expression of OCIAD2 and promoted the expression of DCN. Insulin-like growth factors-1 (IGF-1) inhibited the expression of OCIAD2 and DCN, while fibroblast growth factors 5 (FGF5) promoted the expression of these genes. MT and IGF-1 promoted OCIAD2 synergistically, while MT and FGF5 inhibited the genes simultaneously. MT+IGF-1/MT+FGF5 inhibited DCN gene. RNAi technology showed that OCIAD2 expression was promoted, while that of DCN was inhibited. Conclusion: Activation of bone morphogenetic protein (BMP) signaling pathway up-regulated OCIAD2 expression and stimulated SF to control cell proliferation. DCN gene affected hair follicle morphogenesis and periodic changes by promoting transforming growth $factor-{\beta}$ ($TGF-{\beta}$) and BMP signaling pathways. OCIAD2 and DCN genes have opposite effects on $TGF-{\beta}$ signaling pathway and inhibit each other to affect the hair growth.

• Asian-Australasian Journal of Animal Sciences
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• 제17권11호
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• pp.1496-1500
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• 2004

In the mammalian ovary the follicular fluid contains proteins and peptides which play an important role in growth, development and maturation of oocytes. The gonadotropins and some other factors work synergistically and regulate ovarian functions. In the present study the effect of follicular fluid proteins (FFP) and gonadotropins on progesterone secretion by granulosa cells (GC) from buffalo ovary, was investigated during culture. The follicular fluid was collected from small (<5 mm), and medium (5-8 mm) follicles obtained from buffalo ovaries. The follicular fluid from medium follicles was fractionated with ammonium sulphate at 80% saturation. The precipitated protein fraction was further resolved in to minor (peaks I, III) and major (peak II) proteins using gel filtration (Sephadex G-200). The FFP from small follicles and major FFP (peak II) at a dose of 200 $\mu$g/well, significantly stimulated progesterone secretion by pooled GC (3${\times}10^{5}$ cells/2 ml medium/well). The minor FFP did not show any stimulatory effect. There was a significant increase in progesterone secretion by pooled GC in presence of FFP and LH (10 ng/well), however, FSH (20 ng/well) with FFP exhibited an inhibitory effect. The major FFP and gonadotropins were also studied for their effect on progesterone production by GC isolated from medium and large size follicles. The GC from medium follicles were more responsive to FSH and FFP whereas GC from large follicles exhibited enhanced progesterone secretion with LH and FFP. These results indicated that FFP have their own stimulatory effect and also act synergistically with gonadotropins. The significantly different response shown by GC, for steroid hormone secretion, is based on their stage of growth and differentiation. The purification and characterization of such steroidogenic proteins may help in elucidating their role in growth and differentiation of granulosa cells.

• BMB Reports
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• 제36권6호
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• pp.603-607
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• 2003

Abstract Plant lipid transfer proteins (LTPs) are a class of proteins whose functions are still unknown. Some are proposed to have antimicrobial activities. To understand whether LTP110, a rice LTP that we previously identified from rice leaves, plays a role in the protection function against some serious rice pathogens, we investigated the antifungal and antibacterial properties of LTP110. A cDNA sequence, encoding the mature peptide of LTP110, was cloned into the Impact-CN prokaryotic expression system. The purified protein was used for an in vitro inhibition test against rice pathogens, Pyricularia oryzae and Xanthomonas oryzae. The results showed that LTP110 inhibited the germination of Pyricularia oryzae spores, and its inhibitory activity decreased in the presence of a divalent cation. This suggests that the antifungal activity is affected by ions in the media; LTP110 only slightly inhibited the growth of Xanthomonas oryzae. However, the addition of LTP110 to cultured Chinese hamster ovarian cells did not retard growth, suggesting that the toxicity of LTP110 is only restricted to some cell types. Its antimicrobial activity is potentially due to interactions between LTP and microbe-specific structures.