Document Detail


Mutational analysis of an archaebacterial promoter: essential role of a TATA box for transcription efficiency and start-site selection in vitro.
MedLine Citation:
PMID:  2124695     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
By using a recently developed in vitro transcription assay, the 16S/23S rRNA-encoding DNA promoter from the archaebacterium Sulfolobus sp. B12 was dissected by deletion and linker substitution mutagenesis. The analysis of 5' and 3' deletion mutants defined a core promoter region between positions -38 and -2 containing all information for efficient and specific transcription. Further characterization of this region by linker substitution mutagenesis indicated two sequence elements important for promoter function--one located between positions -38 and -25 (distal promoter element) and the other one located between positions -11 and -2 (proximal promoter element). The distal promoter element encompassed the TATA-like "box A" element located approximately 26 nucleotides upstream of the majority of transcription start sites in archaebacteria (Archaeobacteria). All mutations within this box A motif virtually abolished promoter function. Complete inactivation of the proximal promoter element was dependent on extensive mutagenesis; this element is not conserved between archaebacterial promoters except for a high A + T content in stable RNA gene promoters from Sulfolobus. Mutants containing insertions or deletions between the distal and proximal promoter elements were only slightly affected in their transcription efficiency but displayed a shift in their major initiation site, retaining an essentially fixed distance between the distal promoter element and the transcription start site. Thus, efficient transcription and start-site selection were dependent on a conserved TATA-like sequence centered approximately 26 nucleotides upstream of the initiation site, a situation unlike that of eubacterial promoters but resembling the core structure of most eukaryotic RNA polymerase II (and some RNA polymerase III) promoters. This finding suggests a common evolutionary origin of these promoters consistent with the known similarities between archaebacterial and eukaryotic RNA polymerases.
Authors:
W D Reiter; U Hüdepohl; W Zillig
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Proceedings of the National Academy of Sciences of the United States of America     Volume:  87     ISSN:  0027-8424     ISO Abbreviation:  Proc. Natl. Acad. Sci. U.S.A.     Publication Date:  1990 Dec 
Date Detail:
Created Date:  1991-02-07     Completed Date:  1991-02-07     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  7505876     Medline TA:  Proc Natl Acad Sci U S A     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  9509-13     Citation Subset:  IM    
Affiliation:
Max-Planck-Institut für Biochemie, Martinsried, Federal Republic of Germany.
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MeSH Terms
Descriptor/Qualifier:
Archaea / genetics*
Base Sequence
Chromosome Deletion
DNA, Ribosomal / genetics
Genes, Bacterial*
Molecular Sequence Data
Mutagenesis, Site-Directed*
Plasmids
Promoter Regions, Genetic*
RNA, Ribosomal, 16S / genetics
RNA, Ribosomal, 23S / genetics
TATA Box*
Transcription, Genetic*
Chemical
Reg. No./Substance:
0/DNA, Ribosomal; 0/RNA, Ribosomal, 16S; 0/RNA, Ribosomal, 23S
Comments/Corrections

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