Utilization of MHC class I transgenic mice for development of minigene DNA vaccinesencoding multiple HLA-restricted CTL epitopes.
AuthorsIshioka GY, Fikes J, Hermanson G, Livingston B, Crimi C, Qin M, del Guercio MF, Oseroff C, Dahlberg C, Alexander J, Chesnut RW, Sette A
JournalJ Immunol
PubMed ID10201910
'We engineered a multiepitope DNA minigene encoding nine dominant HLA-A2.1- and A11-restricted epitopes from the polymerase, envelope, and core proteins of hepatitis B virus and HIV, together with the PADRE (pan-DR epitope) universal Th cell epitope and an endoplasmic reticulum-translocating signal sequence. Immunization of HLA transgenic mice with this construct ... More
Interactions between protein kinase CK2 and Pin1. Evidence for phosphorylation-dependent interactions.
Authors Messenger Moira M; Saulnier Ronald B; Gilchrist Andrew D; Diamond Phaedra; Gorbsky Gary J; Litchfield David W;
JournalJ Biol Chem
PubMed ID11940573
'The peptidyl-prolyl isomerase Pin1 interacts in a phosphorylation-dependent manner with several proteins involved in cell cycle events. In this study, we demonstrate that Pin1 interacts with protein kinase CK2, an enzyme that generally exists in tetrameric complexes composed of two catalytic CK2 alpha and/or CK2 alpha'' subunits together with two ... More
Role of CCAA nucleotide repeats in regulation of hemoglobin and hemoglobin-haptoglobin binding protein genes of haemophilus influenzae.
AuthorsRen Z, Jin H, Whitby PW, Morton DJ, Stull TL
JournalJ Bacteriol
PubMed ID10482534
'Haemophilus influenzae utilizes hemoglobin and hemoglobin-haptoglobin as heme sources. The H. influenzae hemoglobin- and hemoglobin-haptoglobin binding protein genes, hgpA, hgpB, and hgpC, contain lengths of tetrameric CCAA repeats. Using an hgpA-lacZ translational gene fusion, we demonstrate phase-variable expression of lacZ associated with alteration in the length of the CCAA repeat ... More
Regulation of the Bub2/Bfa1 GAP complex by Cdc5 and cell cycle checkpoints.
Authors Hu F; Wang Y; Liu D; Li Y; Qin J; Elledge S J;
JournalCell
PubMed ID11733064
'During mitosis, a ras-related GTPase (Tem1) binds GTP and activates a signal transduction pathway to allow mitotic exit. During most of the cell cycle, Tem1 function is antagonized by a GTPase-activating protein complex, Bfa1/Bub2. How the Bfa1/Bub2 complex is regulated is not well understood. We find that Polo/Cdc5 kinase acts ... More
Substantially enhanced cloning efficiency of SAGE (Serial Analysis of Gene Expression) byadding a heating step to the original protocol.
AuthorsKenzelmann M, Muhlemann K
JournalNucleic Acids Res
PubMed ID9889294
'The efficiency of the original SAGE (Serial Analysis of Gene Expression) protocol was limited by a small average size of cloned concatemers. We describe a modification of the technique that overcomes this problem. Ligation of ditags yields concatemers of various sizes. Small concatemers may aggregate and migrate with large ones ... 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
Ets-1 regulates fli-1 expression in endothelial cells. Identification of ETS binding sites in the fli-1 gene promoter.
'To understand the role of the Ets-1 transcription factor during angiogenesis, we have overexpressed it in endothelial cells and analyzed the levels of expression of several candidate target genes involved in angiogenesis. The transcripts levels of the ETS transcription factor fli-1 are specifically up-regulated in endothelial cells, which overexpress Ets-1, ... More
Shielding of the A1 Domain by the D'D3 Domains of von Willebrand Factor Modulates Its Interaction with Platelet Glycoprotein Ib-IX-V.
AuthorsUlrichts H, Udvardy M, Lenting PJ, Pareyn I, Vandeputte N, Vanhoorelbeke K, Deckmyn H,
JournalJ Biol Chem
PubMed ID16373331
Soluble von Willebrand factor (VWF) has a low affinity for platelet glycoprotein (GP) Ibalpha and needs immobilization and/or high shear stress to enable binding of its A1 domain to the receptor. The previously described anti-VWF monoclonal antibody 1C1E7 enhances VWF/GPIbalpha binding and recognizes an epitope in the amino acids 764-1035 ... More
Nonreceptor tyrosine protein kinase pp60c-src in spatial learning: synapse-specific changes in its gene expression, tyrosine phosphorylation, and protein-protein interactions.
AuthorsZhao W, Cavallaro S, Gusev P, Alkon DL
JournalProc Natl Acad Sci U S A
PubMed ID10884433
c-src is a nonreceptor tyrosine protein kinase that is highly concentrated in synaptic regions, including synaptic vesicles and growth cones. Here, we report that the mRNA signal of pp60c-src is widely distributed in the rat brain with particularly high concentrations in the hippocampus. After spatial maze learning, up-regulation of c-src ... More
The salicylate-derived mycobactin siderophores of Mycobacterium tuberculosis are essential for growth in macrophages.
AuthorsDe Voss JJ, Rutter K, Schroeder BG, Su H, Zhu Y, Barry CE 3rd
JournalProc Natl Acad Sci U S A
PubMed ID10655517
Mycobacterium tuberculosis is an important pathogen of mammals that relies on 2-hydroxyphenyloxazoline-containing siderophore molecules called mycobactins for the acquisition of iron in the restrictive environment of the mammalian macrophage. These compounds have been proposed to be biosynthesized through the action of a cluster of genes that include both nonribosomal peptide ... More
BRCA1 interacts with components of the histone deacetylase complex.
AuthorsYarden RI, Brody LC
JournalProc Natl Acad Sci U S A
PubMed ID10220405
Germ-line mutations in the BRCA1 tumor-suppressor gene are associated with an increased susceptibility to breast and ovarian cancer. BRCA1 contains a carboxyl-terminal domain (BRCT) that is shared with several other proteins involved in maintaining genome integrity. In an effort to understand the function of BRCA1, we sought to isolate proteins ... More
Targeted manipulation of maize genes in vivo using chimeric RNA/DNA oligonucleotides
AuthorsZhu T, Peterson DJ, Tagliani L, St Clair G, Baszczynski CL, Bowen B
JournalProc Natl Acad Sci U S A
PubMed ID10411950
Site-specific heritable mutations in maize genes were engineered by introducing chimeric RNA/DNA oligonucleotides. Two independent targets within the endogenous maize acetohydroxyacid synthase gene sequence were modified in a site-specific fashion, thereby conferring resistance to either imidazolinone or sulfonylurea herbicides. Similarly, an engineered green fluorescence protein transgene was site-specifically modified in ... More
Yeast flavin-containing monooxygenase is induced by the unfolded protein response.
AuthorsSuh JK, Robertus JD
JournalProc Natl Acad Sci
PubMed ID10618381
Flavin-containing monooxygenase from yeast (yFMO) carries out the O(2)- and NADPH-dependent oxidation of biological thiols, including oxidizing glutathione to glutathione disulfide. FMO provides a large fraction of the oxidizing necessary for proper folding of disulfide bond-containing proteins; deletion of the enzyme reduces proper folding of endogenous carboxypeptidase Y by about ... More
Initiation of hepatitis delta virus genome replication
AuthorsDingle K, Bichko V, Zuccola H, Hogle J, Taylor J
JournalJ Virol
PubMed ID9573243
The small, 195-amino-acid form of the hepatitis delta virus (HDV) antigen (deltaAg-S) is essential for genome replication, i.e., for the transcription, processing, and accumulation of HDV RNAs. To better understand this requirement, we used purified recombinant deltaAg-S and HDV RNA synthesized in vitro to assemble high-molecular-weight ribonucleoprotein (RNP) structures. After ... More
A lipid-regulated docking site on vinculin for protein kinase C.
Authors Ziegler Wolfgang H; Tigges Ulrich; Zieseniss Anke; Jockusch Brigitte M;
JournalJ Biol Chem
PubMed ID11741957
During cell spreading, binding of actin-organizing proteins to acidic phospholipids and phosphorylation are important for localization and activity of these proteins at nascent cell-matrix adhesion sites. Here, we report on a transient interaction between the lipid-dependent protein kinase Calpha and vinculin, an early component of these sites, during spreading of ... More
Aspartate 313 in the Streptomyces plicatus hexosaminidase plays a critical role in substrate-assisted catalysis by orienting the 2-acetamido group and stabilizing the transition state.
AuthorsWilliams SJ, Mark BL, Vocadlo DJ, James MN, Withers SG
JournalJ Biol Chem
PubMed ID12171933
SpHex, a retaining family 20 glycosidase from Streptomyces plicatus, catalyzes the hydrolysis of N-acetyl-beta-hexosaminides. Accumulating evidence suggests that the hydrolytic mechanism involves substrate-assisted catalysis wherein the 2-acetamido substituent acts as a nucleophile to form an oxazolinium ion intermediate. The role of a conserved aspartate residue (D313) in the active site ... More
Novel baculovirus DNA elements strongly stimulate activities of exogenous and endogenous promoters.
A DNA sequence upstream from the polyhedrin gene of baculovirus Autographa californica nucleopolyhedrovirus (AcMNPV) was found to activate strongly the expression of full or minimal promoters derived from AcMNPV and other sources. Promoters tested included the minimal CMV (CMVm) promoter from human cytomegalovirus, the full heat shock 70 promoter from ... More
Plasma membrane Ca2+ATPase isoform 4b is cleaved and activated by caspase-3 during the early phase of apoptosis.
The plasma membrane Ca(2+) pump (PMCA) is an essential element in the complex of mechanisms that maintain low intracellular Ca(2+) concentration in the living cell. This pump is tightly regulated by calmodulin through binding to a high affinity calmodulin-binding domain at the C terminus that also serves as an autoinhibitor ... More