+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Development of an in vitro transcription system for Neurospora crassa mitochondrial DNA and identification of transcription initiation sites



Development of an in vitro transcription system for Neurospora crassa mitochondrial DNA and identification of transcription initiation sites



Molecular and Cellular Biology 9(9): 3603-3613



We have developed an in vitro transcription system for Neurospora crassa mitochondrial DNA (mtDNA) and used it to identify transcription initiation sites at the 5' ends of the genes encoding the mitochondrial small and large rRNA and cytochrome b (cob). The in vitro transcription start sites correspond to previously mapped 5' ends of major in vivo transcripts of these genes. Sequences around the three transcription initiation sites define a 15-nucleotide consensus sequence, 5'-TTAGARA(T/G)G(T/G)ARTRR-3', all or part of which appears to be an element of an N. crassa mtDNA promoter. A somewhat looser 11-nucleotide consensus sequence, 5'-TTAGARR(T/G)R(T/G)A-3', was derived by including two additional promoters identified recently. Group I extranuclear mutants, such as [poky] and (SG-3], have a 4-base-pair (bp) deletion in the consensus sequence at the 5' end of the mitochondrial small rRNA and are grossly deficient in mitochondrial small rRNA (R. A. Akins and A. M. Lambowitz, Proc. Natl. Acad. Sci. USA 81:3791-3795, 1984). We show here that the 4-bp deletion in the consensus sequence decreases in vitro transcription from this site by more than 99%. N. crassa mtDNA is similar to Saccharomyces cerevisiae mtDNA in having multiple promoters, including separate promoters for the genes encoding the mitochondrial small and large rRNAs. Our results suggests that the primary effect of the 4-bp deletion in group I extranuclear mutants is to inhibit transcription of the mitochondrial small rRNA, leading to severe deficiency of mitochondrial small rRNA and small ribosomal subunits.

Please choose payment method:






(PDF emailed within 0-6 h: $19.90)

Accession: 007198281

Download citation: RISBibTeXText

PMID: 2528684

DOI: 10.1128/mcb.9.9.3603


Related references

Transcription of mitochondrial DNA in vitro from Neurospora crassa. European Journal of Biochemistry 21(4): 478-488, 1971

Characterization of transcription initiation sites on the soybean mitochondrial genome allows identification of a transcription-associated sequence motif. Molecular and General Genetics 228(3): 345-355, 1991

Sequence analysis of wheat mitochondrial transcripts capped in vitro: definitive identification of transcription initiation sites. Current Genetics 20(3): 245-251, 1991

A simple and sensitive method for determining transcription initiation site: identification of two transcription initiation sites in rat group Ii phospholipase A 2 gene. Nucleic Acids Research 18(23): 6997-7002, 1990

A simple and sensitive method for determining transcription initiation site: identification of two transcription initiation sites in rat group II phospholipase A2 gene. Nucleic Acids Research 18(23): 6997-7002, 1990

DNA methylation inhibits elongation but not initiation of transcription in Neurospora crassa. Genes & Development 11(18): 2383-2395, 1997

DNA methylation inhibits elongation but not initiation of transcription in Neurospora crassa. Genes & Development 11(18): 2383-2395, 1997

Identification of initiation sites for transcription of Xenopus laevis mitochondrial DNA. Journal of Biological Chemistry 261(18): 8488-8494, 1986

Physical map of Neurospora crassa mitochondrial DNA and its transcription unit for ribosomal RNA. Nucleic Acids Research 3(11): 3101-3108, 1976

In Vitro Transcription Of Yeast 2 m Dna By Homologous Rna Polymerase Ii. Mapping Of Initiation Sites And Effects Of Protein Factors On The Selectivity Of Transcription. Biochemical Society Transactions 9(2): 169P.4-169P, 1981

Transcription of cloned neurospora crassa genes in vitro. Journal of Cellular Biochemistry Suppl. (10 Part D): 162, 1986

Characterization of primary transcripts and identification of transcription initiation sites on the heavy and light strands of mouse mitochondrial DNA. Biochemistry 28(2): 763-769, 1989

In vitro transcription of yeast 2 micron dna by homologous rna polymerase ii mapping of initiation sites and effects of protein factors on the selectivity of transcription. Biochemical Society Transactions 9(2): 169P, 1981

In vitro transcription of the dispersed 5s ribosomal rna genes of neurospora crassa. Journal of Cellular Biochemistry Suppl. (7 Part A): 113, 1983

Differential transcription of nonrepeated dna during development of neurospora crassa. Developmental Biology 43(1): 35-41, 1975