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PMID: 3405223 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Template sequences required for transcription of Xenopus laevis mitochondrial DNA from two bidirectional promoters.

Molecular and cellular biology ·Vol. 8 ·No. 7 ·1988-07-00 ·Pages 2917-24

Bogenhagen DF, Romanelli MF

Abstract

Previous work from our laboratory has shown that transcription of Xenopus laevis mitochondrial DNA initiates both in vivo and in vitro from bidirectional promoters located between the gene for tRNA(Phe) and the 5' termini of displacement loop DNA strands. A consensus sequence matching the octanucleotide ACGTTATA surrounds each transcription start site. In the present study, we used in vitro mutagenesis to define sequences required for specific transcription in vitro. First, cloned mitochondrial DNA templates generated by deletion mutagenesis were transcribed in vitro to define the limits of functional promoters. The bidirectional promoter located approximately 33 nucleotides upstream from the gene for tRNA(Phe), termed promoter 1, was studied in greatest detail. The results confirmed the hypothesis that the consensus octanucleotide sequence surrounding each start site is an essential promoter element. A duplex 18-base-pair oligonucleotide encoding the symmetrical promoter 1 region was synthesized and cloned in a plasmid vector. This synthetic oligonucleotide was sufficient to support bidirectional transcription. Point mutations within this oligonucleotide were used to identify critical residues within the consensus sequence.

MeSH Terms
Animals Base Sequence Chromosome Deletion DNA, Mitochondrial/metabolism Molecular Sequence Data Mutation Promoter Regions, Genetic Regulatory Sequences, Nucleic Acid Transcription, Genetic Xenopus laevis
Chemicals
DNA, Mitochondrial
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Bogenhagen D F
Department of Pharmacological Sciences, State University of New York, Stony Brook 11794.
Romanelli M F
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38 references, click to expand
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.8.7.2917-2924.1988
Published
1988-07-00
Pages
2917-24
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC363511
Subset
IM
External Links
PubMed source DOI: 10.1128/mcb.8.7.2917-2924.1988 PMC Full Text View journal details
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