An iron-regulated gene, magA, encoding an iron transport protein of Magnetospirillum sp. strain AMB-1.
AuthorsNakamura C, Burgess JG, Sode K, Matsunaga T
JournalJ Biol Chem
PubMed ID7499342
Magnetospirillum sp. AMB-1 is a freshwater magnetic bacterium which synthesizes intracellular particles of magnetite (Fe3O4). A genomic DNA fragment required for synthesis of magnetic particles was previously isolated from a nonmagnetic transposon Tn5 mutant. We have determined the complete nucleotide sequence of this fragment. The 2975-base pair region contains two ... More
Activation of cAMP-responsive genes by stimuli that produce long-term facilitation in Aplysia sensory neurons.
AuthorsKaang BK, Kandel ER, Grant SG
JournalNeuron
PubMed ID8384857
'One of the hallmarks of long-term memory in both vertebrates and invertebrates is the requirement for new protein synthesis. In sensitization of the gill-withdrawal reflex in Aplysia, this requirement can be studied on the cellular level. Here, long-term but not short-term facilitation of the monosynaptic connections between the sensory and ... More
A HPLC-based chloramphenicol acetyltransferase assay for assessing hair growth: comparison of the sensitivity of UV and fluorescence detection.
AuthorsWaldon DJ, Kubicek MF, Johnson GA, Buhl AE
JournalEur J Clin Chem Clin Biochem
PubMed ID7679931
'In our attempt to measure hair growth by hair-specific markers, we used transgenic mice to express the chloramphenicol acetyltransferase gene under the control of an ultrahigh sulphur keratin gene promoter. To quantitate expression of the keratin gene, we required a chloramphenicol acetyltransferase assay which could measure enzyme activity in a ... More
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
Regulation of prodynorphin gene expression in the ovary: distal DNA regulatory elements confer gonadotropin regulation of promoter activity.
'Examination of the regulation of prodynorphin (pro-DYN) promoter activity is limited by the absence of a good cell model. The discovery of pro-DYN mRNA and derived peptides in the reproductive tract led us to examine the cellular localization and hormonal regulation of ovarian pro-DYN expression and to evaluate normal granulosa ... More
Use of fluorescent chloramphenicol derivative as a substrate for chloramphenicol acetyltransferase assays.
AuthorsHruby DE, Wilson EM
JournalMethods Enzymol
PubMed ID1479909
Use of a fluorescent chloramphenicol derivative as a substrate for CAT assays.
AuthorsHruby DE, Brinkley JM, Kang HC, Haugland RP, Young SL, Melner MH
JournalBiotechniques
PubMed ID2317370
Detecting enzymatic activity in cells using fluorogenic substrates.
AuthorsHaugland RP
JournalBiotech Histochem
PubMed ID8580208
Fluorogenic substrates can detect enzymatic activity associated with cells. It is difficult, however, to detect activity within a single cell or in an organelle since hydrolytic substrates yield products that rapidly leak from the cell. Several new solutions are presented including trapping the fluorescent product in membranes, in cell organelles, ... 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
CHOP/GADD153 gene expression response to cellular stresses inhibited by prior exposure to ultraviolet light wavelength band C (UVC). Inhibitory sequence mediating the UVC response localized to exon 1.
AuthorsSchmitt-Ney M, Habener JF
JournalJ Biol Chem
PubMed ID11010973
CHOP/GADD153 is both an activating and repressing transcription factor that is markedly induced in response to a variety of cellular stresses. The CHOP/GADD153 gene was originally cloned because of its inducibility by ultraviolet light wavelength band C (UVC) and has since been found to be activated in response to many ... More
A nonradioactive assay for transfected chloramphenicol acetyltransferase activity using fluorescent substrates.
AuthorsYoung SL, Barbera L, Kaynard AH, Haugland RP, Kang HC, Brinkley M, Melner MH
JournalAnal Biochem
PubMed ID1785695
Studies of the transcriptional activity of gene promoters have been greatly assisted by the widespread use of the chloramphenicol acetyltransferase (CAT) gene as a reporter gene. Previous techniques for assaying CAT enzymatic activity have utilized radioactive substrates or cofactors with the resulting complications of handling radioactive materials. We report here ... 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