Abstract
Plant mitochondrial promoters are poorly conserved but generally share a loose consensus sequence spanning approximately 17 nucleotides. Using a homologous in vitro transcription system, we have previously shown that an 11-nucleotide sequence within this region comprises at least part of the maize mitochondrial atp1 promoter (W. Rapp and D. Stern, EMBO J. 11:1065-1073, 1992). We have extended this finding by using a series of linker-scanning and point mutations to define the atp1 promoter in detail. Our results show that mutations at positions -12 to +5, relative to the major transcription start site, can decrease initiation rates to between < 10 and 40% of wild-type levels. Some mutations, scattered throughout this region, have lesser effects or no effect. Taken together, our data suggest a model in which the atp1 promoter consists of a central domain extending from -7 to +5 and an upstream domain of 1 to 3 bp that is centered around -11 to -12. Because many mutations within this promoter region are tolerated in vitro, the maize atp1 promoter is distinct from the highly conserved yeast mitochondrial promoters.
Full text
PDF![7232](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4d9f/364793/5d7518d9fc69/molcellb00024-0044.png)
![7233](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4d9f/364793/2b7d1f48a296/molcellb00024-0045.png)
![7234](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4d9f/364793/3b5fa8f5e5dc/molcellb00024-0046.png)
![7235](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4d9f/364793/e2c6d1f7e8bd/molcellb00024-0047.png)
![7236](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4d9f/364793/8b6f4513a678/molcellb00024-0048.png)
![7237](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4d9f/364793/d40ecdd74ccc/molcellb00024-0049.png)
![7238](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4d9f/364793/fc1e0d26fb07/molcellb00024-0050.png)
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Biswas T. K., Getz G. S. Regulation of transcriptional initiation in yeast mitochondria. J Biol Chem. 1990 Nov 5;265(31):19053–19059. [PubMed] [Google Scholar]
- Biswas T. K., Ticho B., Getz G. S. In vitro characterization of the yeast mitochondrial promoter using single-base substitution mutants. J Biol Chem. 1987 Oct 5;262(28):13690–13696. [PubMed] [Google Scholar]
- Brown G. G., Auchincloss A. H., Covello P. S., Gray M. W., Menassa R., Singh M. Characterization of transcription initiation sites on the soybean mitochondrial genome allows identification of a transcription-associated sequence motif. Mol Gen Genet. 1991 Sep;228(3):345–355. doi: 10.1007/BF00260626. [DOI] [PubMed] [Google Scholar]
- Clayton D. A. Replication and transcription of vertebrate mitochondrial DNA. Annu Rev Cell Biol. 1991;7:453–478. doi: 10.1146/annurev.cb.07.110191.002321. [DOI] [PubMed] [Google Scholar]
- Covello P. S., Gray M. W. Sequence analysis of wheat mitochondrial transcripts capped in vitro: definitive identification of transcription initiation sites. Curr Genet. 1991 Aug;20(3):245–251. doi: 10.1007/BF00326239. [DOI] [PubMed] [Google Scholar]
- Edwards J. C., Levens D., Rabinowitz M. Analysis of transcriptional initiation of yeast mitochondrial DNA in a homologous in vitro transcription system. Cell. 1982 Dec;31(2 Pt 1):337–346. doi: 10.1016/0092-8674(82)90127-1. [DOI] [PubMed] [Google Scholar]
- Finnegan P. M., Brown G. G. Transcriptional and Post-Transcriptional Regulation of RNA Levels in Maize Mitochondria. Plant Cell. 1990 Jan;2(1):71–83. doi: 10.1105/tpc.2.1.71. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Greenberg B. M., Narita J. O., DeLuca-Flaherty C., Gruissem W., Rushlow K. A., Hallick R. B. Evidence for two RNA polymerase activities in Euglena gracilis chloroplasts. J Biol Chem. 1984 Dec 10;259(23):14880–14887. [PubMed] [Google Scholar]
- Hanic-Joyce P. J., Gray M. W. Accurate transcription of a plant mitochondrial gene in vitro. Mol Cell Biol. 1991 Apr;11(4):2035–2039. doi: 10.1128/mcb.11.4.2035. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jang S. H., Jaehning J. A. The yeast mitochondrial RNA polymerase specificity factor, MTF1, is similar to bacterial sigma factors. J Biol Chem. 1991 Nov 25;266(33):22671–22677. [PubMed] [Google Scholar]
- Masters B. S., Stohl L. L., Clayton D. A. Yeast mitochondrial RNA polymerase is homologous to those encoded by bacteriophages T3 and T7. Cell. 1987 Oct 9;51(1):89–99. doi: 10.1016/0092-8674(87)90013-4. [DOI] [PubMed] [Google Scholar]
- Mulligan R. M., Lau G. T., Walbot V. Numerous transcription initiation sites exist for the maize mitochondrial genes for subunit 9 of the ATP synthase and subunit 3 of cytochrome oxidase. Proc Natl Acad Sci U S A. 1988 Nov;85(21):7998–8002. doi: 10.1073/pnas.85.21.7998. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mulligan R. M., Leon P., Walbot V. Transcriptional and posttranscriptional regulation of maize mitochondrial gene expression. Mol Cell Biol. 1991 Jan;11(1):533–543. doi: 10.1128/mcb.11.1.533. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rapp W. D., Stern D. B. A conserved 11 nucleotide sequence contains an essential promoter element of the maize mitochondrial atp1 gene. EMBO J. 1992 Mar;11(3):1065–1073. doi: 10.1002/j.1460-2075.1992.tb05145.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Reiss T., Link G. Characterization of transcriptionally active DNA-protein complexes from chloroplasts and etioplasts of mustard (Sinapis alba L.). Eur J Biochem. 1985 Apr 15;148(2):207–212. doi: 10.1111/j.1432-1033.1985.tb08826.x. [DOI] [PubMed] [Google Scholar]
- Rushlow K. E., Orozco E. M., Jr, Lipper C., Hallick R. B. Selective in vitro transcription of Euglena chloroplast ribosomal RNA genes by a transcriptionally active chromosome. J Biol Chem. 1980 Apr 25;255(8):3786–3792. [PubMed] [Google Scholar]
- Sawadogo M., Roeder R. G. Factors involved in specific transcription by human RNA polymerase II: analysis by a rapid and quantitative in vitro assay. Proc Natl Acad Sci U S A. 1985 Jul;82(13):4394–4398. doi: 10.1073/pnas.82.13.4394. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schinkel A. H., Koerkamp M. J., Touw E. P., Tabak H. F. Specificity factor of yeast mitochondrial RNA polymerase. Purification and interaction with core RNA polymerase. J Biol Chem. 1987 Sep 15;262(26):12785–12791. [PubMed] [Google Scholar]
- Stern D. B., Newton K. J. Isolation of plant mitochondrial RNA. Methods Enzymol. 1986;118:488–496. doi: 10.1016/0076-6879(86)18095-5. [DOI] [PubMed] [Google Scholar]
- Stern D. B., Newton K. J. Mitochondrial gene expression in Cucurbitaceae: conserved and variable features. Curr Genet. 1985;9(5):395–404. doi: 10.1007/BF00421611. [DOI] [PubMed] [Google Scholar]