Document Detail


Further characterization of a paramyxovirus transcription initiation signal: search for required nucleotides upstream and importance of the N phase context.
MedLine Citation:
PMID:  17412986     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Paramyxovirus genomes contain a linear array of five to ten genes sequentially transcribed by the viral RNA polymerase. mRNA synthesis initiates at a nucleotide signal (gs1) within the genomic promoter located at the genome 3' end. To gain information about the mechanism involved in transcription initiation, a search was carried out for upstream nucleotides required for gs1 and the effects of the gs1 nucleocapsid protein (N) phase context on transcription regulation were determined. For both purposes, tandem promoter mini-genomes carrying a transcription signal ectopically positioned downstream of a replication-only signal were used. The requirement for hygromycin resistance gene expression was used in an attempt to select essential nucleotides within randomized stretches of nucleotides. Nucleotide insertions or deletions were also made on either side of the transcription signal to change its original N phase context in the five remaining possibilities and GFP expression from these modified signals was assessed. Cell cultures resistant to hygromycin treatment were readily obtained following amplification of mini-genomes harbouring randomized sequences. However, selected nucleotides upstream of gs1 could not be identified under conditions where nucleotides within gs1 were selected. In contrast, it was observed that changing the gs1 N phase context progressively decreased transcription by five- to tenfold. These results are discussed in relation to two different mechanisms of transcription initiation.
Authors:
Samuel Cordey; Laurent Roux
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The Journal of general virology     Volume:  88     ISSN:  0022-1317     ISO Abbreviation:  J. Gen. Virol.     Publication Date:  2007 May 
Date Detail:
Created Date:  2007-04-06     Completed Date:  2007-07-24     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  0077340     Medline TA:  J Gen Virol     Country:  England    
Other Details:
Languages:  eng     Pagination:  1555-64     Citation Subset:  IM    
Affiliation:
Department of Microbiology and Molecular Medicine, University of Geneva Medical School, CMU, 1 rue Michel-Servet, CH-1211 Geneva 4, Switzerland.
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MeSH Terms
Descriptor/Qualifier:
Animals
Base Sequence
Cell Line
Genome, Viral
Molecular Sequence Data
Nucleocapsid / genetics
Paramyxovirinae / genetics*
Plasmids
Promoter Regions, Genetic
RNA, Messenger / genetics*
RNA, Viral / genetics*
Reverse Transcriptase Polymerase Chain Reaction
Transcription, Genetic*
Chemical
Reg. No./Substance:
0/RNA, Messenger; 0/RNA, Viral

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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