Improvement in sensitivity of allele-specific PCR facilitates reliable noninvasive
prenatal detection of cystic fibrosis.
AuthorsNasis O, Thompson S, Hong T, Sherwood M, Radcliffe S, Jackson L, Otevrel T,
JournalClin Chem
PubMed ID14764639
BACKGROUND: Cell-free fetal DNA circulating in maternal blood has potential
as a safer alternative to invasive methods of prenatal testing for paternally inherited genetic alterations,
such as cystic fibrosis (CF) mutations. METHODS: We used allele-specific PCR to detect mutated CF D1152H
DNA in the presence of an excess of the corresponding wild-type sequence. ... More
Cooperation of yeast peroxiredoxins Tsa1p and Tsa2p in the cellular defense against oxidative and nitrosative stress.
Authors Wong Chi-Ming; Zhou Yuan; Ng Raymond W M; Kung Hf Hsiang-fu; Jin Dong-Yan;
JournalJ Biol Chem
PubMed ID11741925
'Peroxiredoxins are a family of antioxidant enzymes conserved from bacteria to humans. In Saccharomyces cerevisiae, there exist five peroxiredoxins, among which Tsa2p shares striking homology with the well described Tsa1p but has not been extensively studied. Here we report on the functional characterization of yeast tsa2Delta mutants and the comparison ... More
Phosphorylation regulates the subcellular location and activity of the snail transcriptional repressor.
AuthorsDomÃnguez D, Montserrat-SentÃs B, Virgós-Soler A, Guaita S, Grueso J, Porta M, Puig I, Baulida J, Francà C, GarcÃa de Herreros A,
JournalMol Cell Biol
PubMed ID12832491
'The Snail gene product is a transcriptional repressor of E-cadherin expression and an inducer of the epithelial-to-mesenchymal transition in several epithelial tumor cell lines. This report presents data indicating that Snail function is controlled by its intracellular location. The cytosolic distribution of Snail depended on export from the nucleus by ... More
Molecular cloning and identification of 3'-phosphoadenosine 5'-phosphosulfate transporter.
AuthorsKamiyama S, Suda T, Ueda R, Suzuki M, Okubo R, Kikuchi N, Chiba Y, Goto S, Toyoda H, Saigo K, Watanabe M, Narimatsu H, Jigami Y, Nishihara S,
JournalJ Biol Chem
PubMed ID12716889
'Nucleotide sulfate, namely 3''-phosphoadenosine 5''-phosphosulfate (PAPS), is a universal sulfuryl donor for sulfation. Although a specific PAPS transporter is present in Golgi membrane, no study has reported the corresponding gene. We have identified a novel human gene encoding a PAPS transporter, which we have named PAPST1, and the Drosophila melanogaster ... More
Fragmin60 encodes an actin-binding protein with a C2 domain and controls actin Thr-203 phosphorylation in Physarum plasmodia and sclerotia.
AuthorsSklyarova T, De Corte V, Meerschaert K, Devriendt L, Vanloo B, Bailey J, Cook LJ, Goethals M, Van Damme J, Puype M, Vandekerckhove J, Gettemans J.
JournalJ Biol Chem
PubMed ID12167630
'We report the isolation of a cDNA clone encoding a 60-kDa protein termed fragmin60 that cross-reacts with fragmin antibodies. Unlike other gelsolin-related proteins, fragmin60 contains a unique N-terminal domain that shows similarity with C2 domains of aczonin, protein kinase C, and synaptotagmins. The fragmin60 C2 domain binds three calcium ions, ... More
Cloning and characterization of a lysosomal phospholipase A2, 1-o-acylceramide synthase.
Authors Hiraoka Miki; Abe Akira; Shayman James A;
JournalJ Biol Chem
PubMed ID11790796
'Recently, a novel enzyme, 1-O-acylceramide synthase (ACS), was purified and characterized from bovine brain. This enzyme has both calcium-independent phospholipase A(2) and transacylase activities. The discovery of this enzyme led us to propose a new pathway for ceramide metabolism in which the sn-2-acyl group of either phosphatidylethanolamine or phosphatidylcholine is ... More
Transcriptional Regulation of the Pituitary Vasopressin V1b Receptor Involves a GAGA-binding Protein.
'The role of CT repeats (inverted GAGA box) in the rat vasopressin V1b receptor (V1bR) promoter in the transcriptional regulation of this gene was studied in H32 hypothalamic cells, which express endogenous V1bR. Transfection of a 2.5-kb V1bR fragment (2161 bp upstream and 377 bp downstream of the proximal transcriptional ... More
A CDC45 homolog in Arabidopsis is essential for meiosis, as shown by RNA interference-induced gene silencing.
'CDC45 is required for the initiation of DNA replication in yeast and cell proliferation in mammals and functions as a DNA polymerase alpha loading factor in Xenopus. We have cloned a CDC45 homolog from Arabidopsis whose expression is upregulated at the G1/S transition and in young meiotic flower buds. One-third ... More
Cloning and functional characterization of HDAC11, a novel member of the human histone deacetylase family.
Authors Gao Lin; Cueto Maria A; Asselbergs Fred; Atadja Peter;
JournalJ Biol Chem
PubMed ID11948178
'We have cloned and characterized a human cDNA that belongs to the histone deacetylase family, which we designate as HDAC11. The predicted HDAC11 amino acid sequence reveals an open reading frame of 347 residues with a corresponding molecular mass of 39 kDa. Sequence analyses of the putative HDAC11 protein indicate ... More
A Novel Diacylglycerol-lactone Shows Marked Selectivity in Vitro among C1 Domains of Protein Kinase C (PKC) Isoforms {alpha} and {delta} as Well as Selectivity for RasGRP Compared with PKC{alpha}.
AuthorsPu Y, Perry NA, Yang D, Lewin NE, Kedei N, Braun DC, Choi SH, Blumberg PM, Garfield SH, Stone JC, Duan D, Marquez VE,
JournalJ Biol Chem
PubMed ID15923197
'Although multiple natural products are potent ligands for the diacylglycerol binding C1 domain of protein kinase C (PKC), RasGRP, and related targets, the high conservation of C1 domains has impeded the development of selective ligands. We characterized here a diacylglycerol-lactone, 130C037, emerging from a combinatorial chemical synthetic strategy, which showed ... More
Optimization of fusion PCR for in vitro construction of gene knockout fragments.
Authors Wang Hong-Liang; Postier Bradley L; Burnap Robert L;
JournalBiotechniques
PubMed ID12139251
This paper contains a comparison of proofreading polymerases including Platinum Pfx and Turbo Pfu.
Molecular characterization of the starfish inositol 1,4,5-trisphosphate receptor and its role during oocyte maturation and fertilization.
The release of calcium ions (Ca(2+)) from their intracellular stores is essential for the fertilization of oocytes of various species. The calcium pools can be induced to release Ca(2+) via two main types of calcium channel receptor: the inositol 1,4,5-trisphosphate receptor (IP(3)R) and the ryanodine receptor. Starfish oocytes have often ... More
mRNA PCR-based epitope chase method.
AuthorsDoucet JD, Gauchat D, Lapointe R
JournalMethods Mol Biol
PubMed ID23296942
The identification of specific viral and tumor antigen epitopes recognized by CD4(+) or CD8(+) T lymphocytes remains a challenge. Unfortunately, epitope mapping methods are generally costly and time-consuming. This chapter details a polymerase chain reaction (PCR)-based mRNA epitope identification method called mPEC, which is designed to rapidly and precisely identify ... More
Identification of endopeptidase genes from the genomic sequence of Lactobacillus helveticus CNRZ32 and the role of these genes in hydrolysis of model bitter peptides.
AuthorsSridhar VR, Hughes JE, Welker DL, Broadbent JR, Steele JL,
JournalAppl Environ Microbiol
PubMed ID15932998
Genes encoding three putative endopeptidases were identified from a draft-quality genome sequence of Lactobacillus helveticus CNRZ32 and designated pepO3, pepF, and pepE2. The ability of cell extracts from Escherichia coli DH5alpha derivatives expressing CNRZ32 endopeptidases PepE, PepE2, PepF, PepO, PepO2, and PepO3 to hydrolyze the model bitter peptides, beta-casein (beta-CN) ... More
Fetal fibronectin signaling induces matrix metalloproteases and cyclooxygenase-2 (COX-2) in amnion cells and preterm birth in mice.
AuthorsMogami H, Kishore AH, Shi H, Keller PW, Akgul Y, Word RA
JournalJ Biol Chem
PubMed ID23184961
Fetal fibronectin (fFN) in cervical and vaginal secretions has been used as a predictor of preterm delivery. Here, we clarified the pathological function of fFN on cell type-specific matrix metalloproteinases (MMPs) and prostaglandin synthesis in fetal membranes. Treatment of amnion mesenchymal cells with fFN resulted in dramatic increases in MMP-1 ... More
Cloning of MASK, a novel member of the mammalian germinal center kinase III subfamily, with apoptosis-inducing properties.
Authors Dan Ippeita; Ong Shao-En; Watanabe Norinobu M; Blagoev Blagoy; Nielsen Mogens M; Kajikawa Eriko; Kristiansen Troels Z; Mann Matthias; Pandey Akhilesh;
JournalJ Biol Chem
PubMed ID11741893
We have cloned a novel human GCK family kinase that has been designated as MASK (Mst3 and SOK1-related kinase). MASK is widely expressed and encodes a protein of 416 amino acid residues, with an N-terminal kinase domain and a unique C-terminal region. Like other GCK-III subfamily kinases, MASK does not ... More
The ducky mutation in Cacna2d2 results in altered Purkinje cell morphology and is associated with the expression of a truncated alpha 2 delta-2 protein with abnormal function.
AuthorsBrodbeck J, Davies A, Courtney JM, Meir A, Balaguero N, Canti C, Moss FJ, Page KM, Pratt WS, Hunt SP, Barclay J, Rees M, Dolphin AC,
JournalJ Biol Chem
PubMed ID11756448
The mouse mutant ducky, a model for absence epilepsy, is characterized by spike-wave seizures and cerebellar ataxia. A mutation in Cacna2d2, the gene encoding the alpha 2 delta-2 voltage-dependent calcium channel accessory subunit, has been found to underlie the ducky phenotype. The alpha 2 delta-2 mRNA is strongly expressed in ... More
High resolution X-ray structure of galactose mutarotase from Lactococcus lactis.
Authors Thoden James B; Holden Hazel M;
JournalJ Biol Chem
PubMed ID11907040
Galactose mutarotase plays a key role in normal galactose metabolism by catalyzing the interconversion of beta-D-galactose and alpha-D-galactose. Here we describe the three-dimensional architecture of galactose mutarotase from Lactococcus lactis determined to 1.9-A resolution. Each subunit of the dimeric enzyme displays a distinctive beta-sandwich motif. This tertiary structural element was ... More
Evidence for a stabilizer element in the untranslated regions of Drosophila glutathione S-transferase D1 mRNA.
Authors Akgül Bünyamin; Tu Chen-Pei D;
JournalJ Biol Chem
PubMed ID12118001
The neighboring genes gstD1 and gstD21 share 70% sequence identity. gstD1 encodes a 1,1,1-trichloro-2,2-bis-(P-chlorophenyl)ethane dehydrochlorinase; gstD21, a ligandin. Both of their mRNAs are inducible by pentobarbital but otherwise behave very differently. Intact gstD21 mRNA is intrinsically labile, but becomes stabilized when separated from its native untranslated region (UTR). In contrast, ... More
The role of apolipoprotein A-I helix 10 in apolipoprotein-mediated cholesterol efflux via the ATP-binding cassette transporter ABCA1.
Recent studies of Tangier disease have shown that the ATP-binding cassette transporter A1 (ABCA1)/apolipoprotein A-I (apoA-I) interaction is critical for high density lipoprotein particle formation, apoA-I integrity, and proper reverse cholesterol transport. However, the specifics of this interaction are unknown. It has been suggested that amphipathic helices of apoA-I bind ... More
The CcmE protein of the c-type cytochrome biogenesis system: unusual in vitro heme incorporation into apo-CcmE and transfer from holo-CcmE to apocytochrome.
Authors Daltrop Oliver; Stevens Julie M; Higham Christopher W; Ferguson Stuart J;
JournalProc Natl Acad Sci U S A
PubMed ID12119398
Three key steps of cytochrome c biogenesis in many Gram-negative bacteria, the uptake of heme by the heme chaperone CcmE, the covalent attachment of heme to CcmE, and its subsequent release from CcmE to an apocytochrome c, have been achieved in vitro. apo-CcmE from Escherichia coli preferentially bound to ferric, ... More
Homo- and heterotypic fibrillin-1 and -2 interactions constitute the basis for the assembly of microfibrils.
Fibrillin-1 and fibrillin-2 constitute the backbone of extracellular filaments, called microfibrils. Fibrillin assembly involves complex multistep mechanisms to result in a periodical head-to-tail alignment in microfibrils. Impaired assembly potentially plays a role in the molecular pathogenesis of genetic disorders caused by mutations in fibrillin-1 (Marfan syndrome) and fibrillin-2 (congenital contractural ... More