Initiation of adenovirus transcription was analyzed by incubation of isolated nuclei from virus infected cells in the presence of beta-32P GTP or beta-32P ATP. Nucleotide analysis of RNA from nuclei incubated with beta-32P GTP shows that the label is incorporated exclusively into pppGp and ppGp. Under similar incubation conditions, the label from beta-32P ATP was incorporated primarily into the...

Guanosine monophosphates (GMPs) were intercalated into the interlayer space of layered double hydroxides (LDHs) and the molecular arrangement of GMP was controlled in LDHs. The intercalation conditions such as GMP/LDH molar ratio and reaction temperature were systematically adjusted. When the GMP/LDH molar ratio was 1:2, which corresponds to the charge balance between positive LDH sheets and GM...

Current models for nucleotide sugar use in the Golgi apparatus predict a critical role for the lumenal nucleoside diphosphatase. After transfer of sugars to endogenous macromolecular acceptors, the enzyme converts nucleoside diphosphates to nucleoside monophosphates which in turn exit the Golgi lumen in a coupled antiporter reaction, allowing entry of additional nucleotide sugar from the cytoso...

A sensitive, versatile and economical method to extract and quantify cyclic nucleotide monophosphates (cNMPs) using LC-MS/MS, including both 3',5'-cNMPs and 2',3'-cNMPs, in mammalian tissues and cellular systems has been developed. Problems, such as matrix effects from complex biological samples, are addressed and have been optimized. This protocol allows for comparison of multiple cNMPs in the...

The purine metabolism of several genera of parasitic protozoa, is of interest partly due to their sensitivity to purine analogues, and to differences in the enzymes of purine metabolism compared to mammalian systems [l-6]. Trichomonas vaginalis, the causative agent of trichomonal vaginitis, a mild, but very common, sexually-transmitted disease, has not been studied in this respect; there is a g...

The WHO has declared tuberculosis (TB) a global health emergency. Therefore, there is an urgent need to discover and develop new anti-TB drugs. Here we report on a new category of 5-substituted pyrimidine nucleosides as potent inhibitors of Myco-bacterium tuberculosis growth in vitro. A series of 2'-deoxy-, 3'-azido-2',3'-dideoxy-, and 3'-amino-2',3'-dideoxypyrimidine nucleoside analogues beari...

A double-labeling procedure for sequence analysis of nonradioactive polyribonucleotides is detailed, which is based on controlled endonucleolytic degradation of 3'-terminally (3H)-labeled oligonucleotide-(3') dialcohols and 5"-terminal analysis of the partial (3H)-labeled fragments following their separation according to chain length by polyethyleneimine- (PEI-)cellulose TLC and detection by fl...

We previously reported the synthesis of a borononucleotide analogue of thymidine monophosphate and its association towards the formation of a new borono-linked dinucleotide. Here we describe the completion of the set of four 2'-deoxyborononucleotide analogues of natural nucleotide monophosphates, namely the previously unknown dCbn, dGbn and dAbn. These analogues were all prepared from the respe...

1This study was supported by American Cancer Society Grant E587. 'Present address: University of Pennsylvania, School of Veterinary Medicine, New Bolton Center, Kennen Square, Pa. 19348. 3To whom reprint requests should be addressed. Present address: University of Colorado Medical Center, Department of Microbiology, Denver, Colo. 80220. 4The abbreviations used are: FdUrd, 5-fluoro-2-deoxyuridin...

Chemical and enzymatic procedures are described for the synthesis of the monophosphates and cyclic phosphates of the antiviral acyclonucleoside 9-(1,3-dihydroxy-2-propoxymethyl)-guanine (DHPG), its 3-hydroxymethyl-4-hydroxybutyl analogue, the (R)- and (S)-epimers of 9-(3,4-dihydroxybutyl)guanine, and 9-(2,3-dihydroxypropyl)guanine. The structures, and some conformational features, of all the fo...