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目的利用酵母回转实验和免疫共沉淀实验验证SIAHI和TRB3之间的相互作用并探讨其功能相关性。方法将全长形式的TRB3基因和SIAH1基因分别克隆入酵母表达载体pDBLeu和pPC86中,共转化至MaV203酵母感受态细胞,验证其相互作用,然后分别构建至真核表达载体pCMV—Myc和pFLAG—CMV-2中,采用免疫共沉淀实验进行进一步验证。通过体内泛素化实验检测SIAH1对TRB3蛋白稳定性及泛素化修饰的影响。结果通过在酵母细胞中的回转实验和HEK293rr细胞中的免疫共沉淀实验证实了TRB3与SIAH1之间的相互作用。通过体内泛素化实验证实了S1AH1介导了TRB3的泛素化修饰和降解。结论证实了TRB3与SIAH1之间的相互作用并发现SIAH1介导了TRB3的泛素化修饰和降解,为TRB3蛋白的功能研究提供了新的线索。 相似文献
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目的观察人乳腺细胞系及乳腺组织中SIAH2和P-ERK表达的变化,探讨乳腺癌中SIAH2与P-ERK的关联。方法应用免疫组织化学检测140例乳腺石蜡包埋组织中SIAH2和P-ERK的表达,Western blot检测人乳腺细胞系和23例乳腺浸润性导管癌及癌旁正常乳腺组织中SIAH2和P-ERK的蛋白表达情况,人乳腺癌细胞系表达SIAH2-siRNA后,P-ERK蛋白表达情况。结果乳腺癌中SIAH2和P-ERK阳性率与组织学分级相关(P<0.05),SIAH2和P-ERK呈正相关性。人乳腺癌细胞系以及乳腺癌组织中SIAH2和P-ERK蛋白表达量明显高于人乳腺正常上皮细胞系与癌旁正常乳腺组织中SIAH2和P-ERK蛋白表达量(P<0.05);表达SIAH2-siRNA的乳腺癌细胞系与对照组相比,P-ERK蛋白表达明显减少(P<0.05)。结论 SIAH2、P-ERK过表达与乳腺癌的组织学分级有关。在乳腺癌细胞系中抑制SIAH2表达后,P-ERK表达也减少,因此抑制SIAH2可以抑制ERK通路。 相似文献
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从热带假丝酵母(Candiada tropicalis)T25—14经过紫外线和亚硝酸的多次诱变,获得4株产十一烷l,11二羧酸(DC13)较多的突变株,其中最优的NP-159株以20%(V/V)正十三烷(nC13)为碳源摇瓶发酵4天,DC13达80g/L左右。在16L罐上,以30%(V/V)nC13发酵6天,DC13高达139g/L,回收残烃后,对nC13的转化率为80%以上。后处理收率为78.9%,DC13的纯度为95.3%。 相似文献
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Nunghathai Sawasdee Mutita Junking Piengpaga Ngaojanlar Nattakan Sukomon Thawornchai Limjindaporn Varaporn Akkarapatumwong Pa-thai Yenchitsomanus 《Biochemical and biophysical research communications》2010,401(1):85-91
Kidney anion exchanger 1 (kAE1) mediates chloride (Cl−) and bicarbonate (HCO3−) exchange at the basolateral membrane of kidney α-intercalated cells. Impaired trafficking of kAE1 leads to defect of the Cl−/HCO3− exchange at the basolateral membrane and failure of proton (H+) secretion at the apical membrane, causing a kidney disease - distal renal tubular acidosis (dRTA). To gain a better insight into kAE1 trafficking, we searched for proteins physically interacting with the C-terminal region of kAE1 (Ct-kAE1), which contains motifs crucial for intracellular trafficking, by a yeast two-hybrid (Y2H) system. An adaptor-related protein complex 1 μ1A (AP-1 mu1A) subunit was found to interact with Ct-kAE1. The interaction between either Ct-kAE1 or full-length kAE1 and AP-1 mu1A were confirmed in human embryonic kidney (HEK) 293T by co-immunoprecipitation, affinity co-purification, co-localization, yellow fluorescent protein (YFP)-based protein fragment complementation assay (PCA) and GST pull-down assay. The interacting site for AP-1 mu1A on Ct-kAE1 was found to be Y904DEV907, a subset of YXXØ motif. Interestingly, suppression of endogenous AP-1 mu1A in HEK 293T by small interfering RNA (siRNA) decreased membrane localization of kAE1 and increased its intracellular accumulation, suggesting for the first time that AP-1 mu1A is involved in the kAE1 trafficking of kidney α-intercalated cells. 相似文献
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In this issue of Molecular Cell, Hogues et al. (2008) demonstrate a wholesale shift in the key regulatory protein involved in ribosomal protein (RP) synthesis during the evolution of S. cerevisiae and, en passant, raise interesting questions about the relationship between RP genes and telomeres. 相似文献
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Harry R. Davis Jr. Scott W. Altmann 《Biochimica et Biophysica Acta (BBA)/Molecular and Cell Biology of Lipids》2009,1791(7):679-683
Niemann–Pick C1 Like 1 (NPC1L1) has been identified and characterized as an essential protein in the intestinal cholesterol absorption process. NPC1L1 localizes to the brush border membrane of absorptive enterocytes in the small intestine. Intestinal expression of NPC1L1 is down regulated by diets containing high levels of cholesterol. While otherwise phenotypically normal, Npc1l1 null mice exhibit a significant reduction in the intestinal uptake and absorption of cholesterol and phytosterols. Characterization of the NPC1L1 pathway revealed that cholesterol absorption inhibitor ezetimibe specifically binds to an extracellular loop of NPC1L1 and inhibits its sterol transport function. Npc1l1 null mice are resistant to diet-induced hypercholesterolemia, and when crossed with apo E null mice, are completely resistant to the development of atherosclerosis. Intestinal gene expression studies in Npc1l1 null mice indicated that no exogenous cholesterol was entering enterocytes lacking NPC1L1, which resulted in an upregulation of intestinal and hepatic LDL receptor and cholesterol biosynthetic gene expression. Polymorphisms in the human NPC1L1 gene have been found to influence cholesterol absorption and plasma low density lipoprotein levels. Therefore, NPC1L1 is a critical intestinal sterol uptake transporter which influences whole body cholesterol homeostasis. 相似文献
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遗传性青光眼包括两种主要的类型,原发性开角型青光眼(primary open angle glaucoma,POAG)和原发性先天型青光眼(primary congenital glaucoma,PCG).眼前节发育不良(anterior segment dysgenesis,ASD)是眼发育异常的遗传异质性病,与增长的眼内压和青光眼有关,包括Peter's异常、Rieger's异常、无虹膜和虹膜发育不全.CYPIB1基因是PCG的致病基因,也有少数报道是POAG的修饰基因,或是POAG和ASD的致病基因.本文就CYP1B1基因突变与遗传性青光眼和ASD发育不全的关系及其遗传特点作一综述. 相似文献
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DEPDC1(DEP domain containing 1)是一个新的肿瘤相关基因,在多种恶性肿瘤的发生发展进程中起着重要作用。我们前期工作中在鼻咽癌细胞内沉默了DEPDC1的表达,发现抑制细胞增殖并诱发细胞凋亡。本研究旨在探讨沉默DEPDC1表达后,对鼻咽癌细胞HNE-1和CNE-1侵袭迁移能力的影响及其分子机制。结果显示,siRNA介导DEPDC1表达沉默后,细胞侧向运动能力、侵袭及迁移能力显著降低。qRT-PCR及Western印迹检测发现DEPDC1沉默导致EMT上游关键转录因子Twist1及间质细胞标志分子Vimentin表达显著下调。这些研究表明,鼻咽癌细胞中DEPDC1通过调节Twist1等EMT关键分子的表达在细胞侵袭转移过程中起关键作用。推测DEPDC1在鼻咽癌中高表达可能对于促进其侵袭转移具有重要作用,进而促进肿瘤发生发展,但具体分子机制仍有待更深入研究。 相似文献
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The purpose of this study was to characterize two mutator stocks of yeast which were induced and selected on the basis of high spontaneous reversion rates of the suppressible "ochre" nonsense allele lys1-1. In the mutator stock VA-3, a single mutation, designated mut1-1, is responsible for the increase in the reversion rate of the ochre alleles lys1-1 and arg4-17. In stock VA-105, there are two separate mutator mutations. Tetrad analysis data showed these two loci are loosely linked. Based on complementation data, one of these mutations is at the same locus as mut1-1 and designated mut1-2. The second mutator of stock VA-105 was designated mut2-1. All three mutators are recessive. Both mut1-1 and mut1-2 give a high mutation rate for ochre nonsense suppressor (SUP) loci, but not for the ochre nonsense alleles. On the contrary, the mutation rates of the ochre alleles are greatly reduced. With the mutant mut2-1 there were mutations at both the lys1-1 site and its suppressors; mut2-1 is as effective as mut1-2 but not as effective as mut1-1 in inducing reversions of a missense mutant, his1-7. Neither mut1-1, mut1-2 nor mut2-1 were effective in inducing reversions of a putative frameshift mutation, hom3-10, or in inducing forward mutations to canavanine resistance. 相似文献
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