Quantification of mitochondrial DNA using real-time polymerase chain reaction in patients with premature ovarian failure.

Objective: To quantify mitochondrial DNA using real-time PCR in women with premature ovarian failure (POF) and a control group.

Design: Prospective study.

Setting: Genome Research Center for Reproductive Medicine and Infertility, Korea Ministry of Health & Welfare.

Molecular cloning of rHAUSP encoding a deubiquitinating enzyme in rat testis.

The tumor suppressor protein p53 is stabilized by the herpes-virus-associated ubiquitin-specific protease (HAUSP), a deubiquitinating enzyme. We previously isolated and characterized a mouse orthologue of HAUSP, mHAUSP. In this study, we have identified a rat orthologue of HAUSP, rHAUSP, from the rat testis by RT-PCR using primers used for cloning mHAUSP.

Differential expression of manganese superoxide dismutase sequence variants in near isogenic lines of wheat during cold acclimation.

Numerous sequence variants of wheat (Triticum aestivum L.) manganese superoxide dismutase (MnSOD) genes have been found. Quantitative real-time PCR was used to measure the expression levels of three MnSOD genes distinguished by a variable amino acid, and three genes distinguished by sequence variation in the 3' untranslated region (3' UTR), in wheat plants grown at 20 degrees C and cold-acclimated for 1-4 weeks at 2 degrees C.

Differential mRNA stability to endogenous ribonucleases of the coding region and 3' untranslated regions of wheat (Triticum aestivum L.) manganese superoxide dismutase genes.

The sequences of the 3' untranslated region (UTR) of the manganese superoxide dismutase (MnSOD) genes in wheat (Triticum aestivum) were found to be quite variable with different predicted thermostabilities. The degradation rates of the 3' UTR variants and the coding region were measured following exposure to endogenous nucleases. The degradation rates of the 3' UTR variants for 15 min were not significantly different, meaning the degradation rates of the 3' UTR variants were not directly related to the thermostabilities.

HAUSP, a deubiquitinating enzyme for p53, is polyubiquitinated, polyneddylated, and dimerized.

The tumor suppressor protein p53 is ubiquitinated and neddylated by MDM2 and then degraded by 26S proteasome. However, p53 is stabilized by the HAUSP (Herpes-virus-associated ubiquitin-specific protease) deubiquitinating enzyme. In this study, we discovered that rat HAUSP (rHAUSP) is polyubiquitinated, polyneddylated, and dimerized using co-immunoprecipitation assays.

Expression and functional analyses of mHAUSP regulating apoptosis of cervical adenocarcinoma cells.

p53 tumor suppressor protein is stabilized by the herpes-virus-associated ubiquitin-specific protease (HAUSP), a deubiquitinating enzyme. We previously isolated a mouse orthologue of HAUSP, mHAUSP, encoding 1103 amino acids with a molecular weight of approximately 135 kDa containing highly conserved Cys, Asp (I), His, and Asn/Asp (II) domains. In this study, we investigated the temporal and spatial expression of mHAUSP during the early mouse embryonic development.

The WD-40 repeat motif of Lgl tumor suppressor proteins associated with salt tolerance and temperature sensitivity.

We have recently identified mammalian homologues of lethal giant larvae (Lgl) tumor suppressor gene, rat Rgl-1 and bovine Bgl-1, and demonstrated that they can complement yeast double mutants lacking Sop1 and Sop2, yeast homologues of Lgl. These gene products are capable of regulating cellular viability in restrictive salt and temperature environments. Since Lgl family members contain the WD-40 repeat motif, we investigated its cellular functions using mouse homologue Mgl-1 in the absence of Sop1 and Sop2 in yeasts by complementation.

Trophoblast apoptosis is increased in women with evidence of TH1 immunity.

Interferon-gamma secretion from peripheral blood mononuclear cells (PBMCs) of women with unexplained recurrent spontaneous abortions was increased by trophoblast stimulation. Apoptosis was significantly increased in trophoblast cells cultured with the supernatant of PBMCs from women with evidence of T(H)1 immunity to trophoblast stimulation.

Deubiquitinating enzyme USP36 contains the PEST motif and is polyubiquitinated.

The ubiquitin-mediated protein degradation pathway has been emphasized for the regulation of numerous cellular mechanisms and the significance of deubiquitination, mediated by deubiquitinating (DUB) enzymes, has been emerging as an essential regulatory step to control these cellular mechanisms. Previously, we demonstrated a human DUB enzyme, HeLa DUB-1, expressed in human ovarian cancer cells. Here, we report human USP36, which has the extension of the C-terminal region of HeLa DUB-1 and has conserved amino acid domains as previously shown in other DUBs.

Long oligonucleotide microarrays in wheat: evaluation of hybridization signal amplification and an oligonucleotide-design computer script.

A computer script was written in the Perl language to design equal-length long oligonucleotides from DNA sequences. The script allows the user to specify G + C content, melting temperature, self-complementarity, the maximum number of contiguous duplicate bases, whether to start with the first start codon and whether to report reverse complements. Microarrays were fabricated with 95 oligonucleotides (60 mers) representing 41 genes.

Complementation of temperature tolerance by rat Rgl-1 recessive oncogene in the absence of Saccharomyces cerevisiae Sop genes.

It has been demonstrated that homozygous mutations at the L(2)gl locus in Drosophila result in the development of tumor in the presumptive adult optic centers of the larval brain and of the imaginal discs. We previously cloned an L(2)gl homologue, Rgl-1, in the rat brain. In this study, we analyzed the capability of Rgl-1 in recovering temperature tolerance in the absence of Saccharomyces cerevisiae Sop genes, yeast homologues of the Drosophila recessive oncogene Lethal (2) giant larvae.

Murine Asb-17 expression during mouse testis development and spermatogenesis.

In this study we isolated a murine mAsb-17 from mouse testis by RT-PCR using primers designed based on the sequences from the GenBank database. The sequence analysis showed that mAsb-17 encodes a 295 amino acid polypeptide with a molecular weight of approximately 34 kDa containing two ankyrin repeats and one SOCS box. The amino acid sequence of mASB-17 showed 87.

Essential regions of deubiquitinating enzyme activity and enhancer function for DUB-2A expressed in T-lymphocytes.

Novel subfamily members of deubiquitinating enzyme gene, DUB-1 and DUB-2, are known to be immediate-early genes and are specifically expressed in B-lymphocytes and T-lymphocytes, respectively. Recently, another deubiquitinating enzyme gene DUB-2A, expressed also in T-lymphocytes, has been isolated and we, in this study, found that Dub-2A has isopeptidase activity and there are at least 4 conserved amino acids (Cys60, Asp133, His298, and His307) that are required for catalytic activity of the enzyme. Interestingly, the conserved Asp323 was not essential for the catalytic activity.

A novel cysteine protease HeLa DUB-1 responsible for cleaving the ubiquitin in human ovarian cancer cells.

The regulation of ubiquitin-mediated protein degradation is becoming important for a number of cellular processes. Human HeLa DUB-1 cDNA, encoding a novel deubiquitinating enzyme, was isolated from ovarian cancer cells. It has 1,647 bp nucleotides and encodes a 548 amino acid polypeptide with the molecular weight of approximately 61 kDa.

Structural and functional conservation of the lgl recessive oncogenes (Review).

It is becoming gradually evident that the dysregulated cell proliferation and differentiation in neoplasia are associated with aberrant functions of specific genes in the genome. Genes and their cellular functions involved in cell proliferation and differentiation are highly conserved among different species. Therefore, the illustration of evolutionary relationships among gene products in different species can provide structural and functional insights, leading to neoplastic studies in lower animals that can help to understand the mechanisms of neoplastic development in vertebrates including humans.

Aberrant gene expression associated with recurrent pregnancy loss.

Recent studies indicate that a number of factors including chromosomal abnormalities, immunological feto-maternal rejection, hormonal irregulation and anatomical factors are involved in provoking recurrent pregnancy loss (RPL). This indicates that normal cellular regulation of these factors is required for maintaining normal pregnancy. In addition, it is expected that biological processes for maintaining normal pregnancy require a series of differential gene expression.

Differentially up-regulated genes in proliferating porcine neonatal pancreas cells caused by epidermal growth factor.

Pancreatic duct cells are considered to be a major source for beta-cell regeneration or neogenesis. Although epidermal growth factor (EGF) is a well-known important growth factor for pancreas development, the control of pancreatic duct cell growth and differentiation by EGF is poorly understood. In this study, we focused on identifying the genes that were differentially up-regulated in response to EGF stimulation using monolayer cultured porcine neonatal pancreas cells.

Identification and characterization of murine mHAUSP encoding a deubiquitinating enzyme that regulates the status of p53 ubiquitination.

Recently, it was discovered that herpesvirus-associated ubiquitin-specific protease (HAUSP) in human interacts with p53 protein, and removes the ubiquitin from ubiquitinated p53. Thus, human HAUSP stabilizes the status of p53, induces p53-dependent cell growth repression and apoptosis. In this study, we isolated and characterized a mouse orthologue of HAUSP, mHAUSP.

Inhibitory effect of salmosin, a Korean snake venom-derived disintegrin, on the integrin alphav-mediated proliferation of SK-Mel-2 human melanoma cells.

We have investigated the inhibitory effect of salmosin on integrin-mediated human tumour cell proliferation. SK-Mel-2 human melanoma cell adhesion to denatured collagen or vitronectin was found to be significantly and statistically inhibited by salmosin in a dose-dependent manner (P<0.05).

DUB-3, a cytokine-inducible deubiquitinating enzyme that blocks proliferation.

Previous studies have identified the DUB family of cytokine-regulated murine deubiquitinating enzymes, which play a role in the control of cell proliferation and survival. Through data base analyses and cloning, we have identified a human cDNA (DUB-3) that shows significant homology to the known murine DUB family members. Northern blotting has shown expression of this gene in a number of tissues including brain, liver, and muscle, with two transcripts being apparent (1.

Functional and expression analyses of mgl-1, a mouse orthologue of lethal giant larvae recessive oncogene.

We recently isolated a few mammalian homologue of Drosophila lethal giant larvae (lgl) recessive oncogene, suggesting that there is functional conservation among proteins of this family. The comparison of amino acid sequence for mgl-1 with other lgl family members using the clustal method showed that they are highly homologous. Therefore, we investigated the biological function of mgl-1, a mouse orthologue of lgl, in the absence of Saccharomyces cerevisiae Sop1 and Sop2, the yeast homologues of the lgl recessive oncogene.

DUB-1A, a novel deubiquitinating enzyme subfamily member, is polyubiquitinated and cytokine-inducible in B-lymphocytes.

Recently, we isolated the Dub-2A gene, which encodes a novel murine deubiquitinating enzyme subfamily member, from a bacterial artificial chromosome library clone by PCR amplification with degenerate PCR primers for the Dub-2 cDNA (Baek, K.-H., Mondoux, M.

Expression of angiogenesis- and apoptosis-related genes in chorionic villi derived from recurrent pregnancy loss patients.

Angiogenesis takes place during embryogenesis, characterized by the formation of new blood vessels from pre-existing ones. This biological process is also found in the female reproductive system, wound healing, and cancer development. Apoptosis, programmed cell death, is a physiological process in development, tissue homeostasis, and disease.

WD-40 repeat containing rat lethal giant larvae recessive oncogene, but not m-tomosyn, restores the salt sensitivity in Saccharomyces cerevisiae.

Recently, we cloned rgl-1, a putative rat homologue of the Drosophila tumor suppressor oncogene lethal (2) giant larvae (l(2)gl), from the rat brain. Sequence analysis revealed that rgl-1 encodes a 1,036 amino acid polypeptide containing a domain characteristic of WD-40 repeat sequence. It has been suggested that m-tomosyn is expressed in the rat brain, encodes a 1,116 amino acid polypeptide, contains WD-40 repeat sequence, and is homologous to the Drosophila l(2)gl and mouse mgl-1 genes.

Conjugation and deconjugation of ubiquitin regulating the destiny of proteins.

The homeostasis for a number of cellular proteins is regulated by not only phosphorylation and dephosphorylation, but also ubiquitination and deubiquitination. A number of proteins involved in the degradation of polypeptides have been isolated in various eukaryotic organisms from Saccharomyces cerevisiae to human. Recently, several deubiquitinating enzymes, classified into either the Ub C-terminal hydrolase (UCH) or the Ub-specific processing protease (UBP), have been reported.