Quantitative nonradioactive CAT assays using fluorescent BODIPY 1-deoxychloramphenicol substrates.
AuthorsLefevre CK, Singer VL, Kang HC, Haugland RP
JournalBiotechniques
PubMed ID7495564
'We report the development of fluorescent BODIPY 1-deoxychloramphenicol substrates for chloramphenicol acetyltransferase (CAT). These substrates not only simplify and improve quantitation of CAT activity but also extend the linear range of detection. Because the 1-deoxychloramphenicol derivatives have only one acetylation site, the enzyme reaction creates only one product, whereas chloramphenicol ... More
Sequence-dependent oligomerization of the Neu transmembrane domain suggests inhibition of
AuthorsBeevers AJ, Damianoglou A, Oates J, Rodger A, Dixon AM,
JournalBiochemistry
PubMed ID20180588
'Membrane-spanning epidermal growth factor receptor ErbB2 is of key importance in cell division, in which a dimeric complex of the protein is responsible for tyrosine kinase activation following ligand binding. The rat homologue of this receptor (Neu) is prone to a valine to glutamic acid mutation in the transmembrane domain ... More
Esterases in serum-containing growth media counteract chloramphenicol acetyltransferase activity in vitro.
AuthorsSohaskey CD, Barbour AG
JournalAntimicrob Agents Chemother
PubMed ID10049283
'The spirochete Borrelia burgdorferi was unexpectedly found to be as susceptible to diacetyl chloramphenicol, the product of the enzyme chloramphenicol acetyltransferase, as it was to chloramphenicol itself. The susceptibilities of Escherichia coli and Bacillus subtilis, as well as that of B. burgdorferi, to diacetyl chloramphenicol were then assayed in different ... More
Domains of human respiratory syncytial virus P protein essential for homodimerization and for binding to N and NS1 protein.
AuthorsHengst U, Kiefer P
JournalVirus Genes
PubMed ID10949949
In this report we used the two-hybrid technique to test for binding among human respiratory syncytial virus (HRSV) proteins involved in the control of viral replication. Besides the expected positive interactions for the nucleoprotein (N) with itself and the phosphoprotein (P), our results also demonstrated P-P interaction and P-NS1 binding. ... More
Spirochaeta aurantia has diacetyl chloramphenicol esterase activity.
AuthorsSohaskey CD, Barbour AG
JournalJ Bacteriol
PubMed ID10714999
The free-living spirochete Spirochaeta aurantia was nearly as susceptible to diacetyl chloramphenicol, the product of chloramphenicol acetyltransferase, as it was to chloramphenicol itself. This unexpected susceptibility to diacetyl chloramphenicol was wholly or partly the consequence of intrinsic carboxylesterase activity, as indicated by high-performance liquid chromatography, thin-layer chromatography, and microbiological assays. ... More
Key role of a downstream specificity protein 1 site in cell cycle-regulated transcription of the AP endonuclease gene APE1/APEX in NIH3T3 cells.
AuthorsFung H, Bennett RA, Demple B
JournalJ Biol Chem
PubMed ID11555653
Abasic (apurinic/apyrimidinic or AP) sites are a frequent type of DNA damage that threatens genetic stability. The predominant mammalian enzyme initiating repair of AP sites is the Ape1 AP endonuclease (also called Apex or Hap1), which also facilitates DNA binding by several transcription factors (Ref1 activity). We found that expression ... More
Nox1-dependent reactive oxygen generation is regulated by Rac1.
AuthorsCheng G, Diebold BA, Hughes Y, Lambeth JD
JournalJ Biol Chem
PubMed ID16636067
Rac1 has been implicated in the generation of reactive oxygen species (ROS) in several cell types, but the enzymatic origin of the ROS has not been proven. The present studies demonstrate that Nox1, a homolog of the phagocyte NADPH-oxidase component gp91(phox), is activated by Rac1. When Nox1 is co-expressed along ... More
E2F transcriptional activation requires TRRAP and GCN5 cofactors.
AuthorsLang SE, McMahon SB, Cole MD, Hearing P
JournalJ Biol Chem
PubMed ID11418595
The E2F family of transcription factors regulates the temporal transcription of genes involved in cell cycle progression and DNA synthesis. E2F transactivation is antagonized by retinoblastoma protein (pRb), which recruits chromatin-remodeling proteins such as histone deacetylases and SWI.SNF complexes to the promoter to repress transcription. We hypothesized that E2F proteins ... More
Novel role for Sp1 in phorbol ester enhancement of human platelet thromboxane receptor gene expression.
Authors D'Angelo D D; Oliver B G; Davis M G; McCluskey T S; Dorn G W;
JournalJ Biol Chem
PubMed ID8702673
Expression of platelet thromboxane receptors is transcriptionally increased during megakaryocytic differentiation stimulated by phorbol 12-myristate 13-acetate (PMA). We previously cloned and characterized the promoter region of the human thromboxane receptor gene and localized PMA-responsive elements to a region between 1.84 and 1.95 kilobase pairs (kb) 5' of the transcription initiation ... More
Elk-1, C/EBPalpha, and Pit-1 confer an insulin-responsive phenotype on prolactin promoter expression in Chinese hamster ovary cells and define the factors required for insulin-increased transcription.
AuthorsJacob KK, Stanley FM
JournalJ Biol Chem
PubMed ID11340077
The transcription factor(s) that mediate insulin-increased gene transcription are not well defined. These studies use phenotypic conversion of Rat2 and Chinese hamster ovary (CHO) cells with transcription factors to identify components required for regulation of prolactin promoter activity and its control by insulin. The pituitary-derived GH4 cells contain all of ... More