Localization of transcription initiation sites on the mouse mitochondrial genome
The authors have identified the primary transcription initiation sites on the H and L strands of mouse mitochondrial (mt) genome by mapping the 5' ends of in vitro capped mt RNA, and 5' end labelling of the nascent RNA synthesized in an in vitro mt system. RNA capped with TSP GTP resolve into 4 major (25 to 150 nucleotides) and one minor (0.75 kb) bands on denaturing gels. Only the 25 nucleotide long capped RNA hybridizes to the H strand of D-loop DNA and the rest hybridize to the L-strand DNA probes. S1 protection of capped RNA and DNA hybrids, and primer extention analysis using defined DNA primers show that all of the L-strand specific primary transcripts have a common 5' end mapping at about nucleotide 16,180 +/- 5 of the genome. The 3' ends of the small RNA species map near the start of conserved sequence boxes. The 3' end of the 0.75 Kb RNA maps to the start of gene coding for tRNA/sup Phe/. The 5' end of the capped RNA hybridizing to the H strand maps at about nucleotide 16,275 to 16,280 of the genome indicating a major H strand transcription initiation at this region. The authors have also used an in vitro transcription system which involves the use of mt extract from Ehrlich ascites cells to study transcription initiation. Nascent RNA 5' end labeled with elTSP ATP and GTP closely resemble the electrophoretic pattern and S1 protection pattern obtained with the capped RNA.
- Research Organization:
- Univ. of Pennsylvania, Philadelphia
- OSTI ID:
- 6295547
- Report Number(s):
- CONF-870644-; TRN: 87-028773
- Journal Information:
- Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Vol. 46:6; Conference: 78. annual meeting of the American Society of Biological Chemists conference, Philadelphia, PA, USA, 7 Jun 1987
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
RNA
BIOSYNTHESIS
GENETIC MAPPING
HYBRIDIZATION
BIOLOGICAL LOCALIZATION
DNA
EHRLICH ASCITES TUMOR
ELECTROPHORESIS
GUANYLIC ACID
MICE
MITOCHONDRIA
PHOSPHORUS 32
TRACER TECHNIQUES
TUMOR CELLS
ANIMAL CELLS
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CELL CONSTITUENTS
DAYS LIVING RADIOISOTOPES
EXPERIMENTAL NEOPLASMS
ISOTOPE APPLICATIONS
ISOTOPES
LIGHT NUCLEI
MAMMALS
MAPPING
NUCLEI
NUCLEIC ACIDS
NUCLEOTIDES
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
ORGANOIDS
PHOSPHORUS ISOTOPES
RADIOISOTOPES
RODENTS
SYNTHESIS
VERTEBRATES
550401* - Genetics- Tracer Techniques