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

Journal Article · Fri Jul 01 04:00:00 EDT 1988 · Mol. Cell. Biol.; (United States)

DOI:https://doi.org/10.1128/MCB.8.7.2917· OSTI ID:6272712

Bogenhagen, D F; Romanelli, M F

Previous work from the authors' 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/sup 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, the authors 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/sup 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.

• Research Organization: Dept. of Pharmacological Sciences, State Univ. of New York at Stony Brook, Stony Brook, NY (US)
• OSTI ID: 6272712
• Journal Information: Mol. Cell. Biol.; (United States), Journal Name: Mol. Cell. Biol.; (United States) Vol. 8:7; ISSN MCEBD
• Country of Publication: United States
• Language: English

Related Subjects

550200* -- Biochemistry
59 BASIC BIOLOGICAL SCIENCES
AMPHIBIANS
ANIMALS
AQUATIC ORGANISMS
CELL CONSTITUENTS
CLONING
DNA HYBRIDIZATION
DNA SEQUENCING
DNA-CLONING
FROGS
GENE OPERONS
GENE REPRESSORS
GENES
HYBRIDIZATION
IN VITRO
IN VIVO
MITOCHONDRIA
MOLECULAR STRUCTURE
NUCLEIC ACIDS
NUCLEOPROTEINS
OLIGONUCLEOTIDES
ORGANIC COMPOUNDS
ORGANOIDS
PLASMIDS
PROTEINS
REPLICONS
RNA
STRUCTURAL CHEMICAL ANALYSIS
TRANSCRIPTION
TRANSFER RNA
VERTEBRATES