« Previous AbstractSynthetic genetic interactions with temperature-sensitive clathrin in Saccharomyces cerevisiae. Roles for synaptojanin-like Inp53p and dynamin-related Vps1p in clathrin-dependent protein sorting at the trans-Golgi network    Next AbstractPheromone cCF10 and plasmid pCF10-encoded regulatory molecules act post-transcriptionally to activate expression of downstream conjugation functions »

Proc Natl Acad Sci U S A

Title:		Sensitive detection of bacterial transcription initiation sites and differentiation from RNA processing sites in the pheromone-induced plasmid transfer system of Enterococcus faecalis
Author(s):		Bensing BA; Meyer BJ; Dunny GM;
Address:		"Department of Microbiology, University of Minnesota, Minneapolis 55455,USA"
Journal Title:		Proc Natl Acad Sci U S A
Year:		1996
Volume:		93
Issue:		15
Page Number:		7794 - 7799
DOI:		10.1073/pnas.93.15.7794
ISSN/ISBN:		0027-8424 (Print) 1091-6490 (Electronic) 0027-8424 (Linking)
Abstract:		"A method was developed to detect 5' ends of bacterial RNAs expressed at low levels and to differentiate newly initiated transcripts from processed transcripts produced in vivo. The procedure involves use of RNA ligase to link a specific oligoribonucleotide to the 5' ends of cellular RNAs, followed by production of cDNA and amplification of the gene of interest by PCR. The method was used to identify the precise sites of transcription initiation within a 10-kb region of the pheromone-inducible conjugative plasmid pCF10 of Enterococcus faecalis. Results confirmed the 5' end of a very abundant, constitutively produced transcript (from prgQ) that had been mapped previously by primer extension and defined the initiation point of a less abundant, divergently transcribed message (from prgX). The method also showed that the 5' end of a pheromone-inducible transcript (prgB) that had been mapped by primer extension was generated by processing rather than new initiation. In addition, the results provided evidence for two promoters, 3 and 5 kb upstream of prgB, and indicated that only the transcripts originating 5 kb upstream may be capable of extending to prgB"
Keywords:		"Amino Acid Sequence Base Sequence Binding Sites Conjugation, Genetic DNA Primers DNA-Directed RNA Polymerases/metabolism Enterococcus faecalis/*genetics/*metabolism Genes, Bacterial Molecular Sequence Data Oligodeoxyribonucleotides Pheromones/*physiology;"
Notes:		"MedlineBensing, B A Meyer, B J Dunny, G M eng 1T32GM08347/GM/NIGMS NIH HHS/ R01GM49530/GM/NIGMS NIH HHS/ Research Support, U.S. Gov't, P.H.S. 1996/07/23 Proc Natl Acad Sci U S A. 1996 Jul 23; 93(15):7794-9. doi: 10.1073/pnas.93.15.7794"