+ 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

Precise mapping and characterization of the RNA primers of DNA replication for a yeast hypersuppressive petite by in vitro capping with guanylyltransferase



Precise mapping and characterization of the RNA primers of DNA replication for a yeast hypersuppressive petite by in vitro capping with guanylyltransferase



Nucleic Acids Research 26(5): 1309-1316



The active origins of DNA replication for yeast (Saccharomyces cerevisiae) mitochondrial DNA share 280 conserved base pairs and have a promoter. Since intact replication intermediates retain their initiating ribonucleotide triphosphate, we used guanylyl-transferase to in vitro cap the replication intermediates present in restriction enzyme-cut DNA from an ori-5 hypersuppressive petite. Restriction mapping and RNA sequencing of these labeled intermediates showed that each DNA strand is primed at a single discrete nucleotide, that one primer starts at the promoter and that the other primer starts 34 nt away, outside the conserved region. Deoxyribonuclease digestion of the capped fragments left resistant RNA primers, which enabled identification of zones of transition from RNA to DNA synthesis. Some of the results contradict the prevailing model for priming at the yeast mitochondrial origins.

Please choose payment method:






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

Accession: 009225584

Download citation: RISBibTeXText

PMID: 9469842

DOI: 10.1093/nar/26.5.1309


Related references

Replication and preferential inheritance of hypersuppressive petite mitochondrial DNA. Embo Journal 20(7): 1807-1817, 2001

Identification of multiple transcriptional initiation sites on the yeast mitochondrial genome by in vitro capping with guanylyltransferase. Journal of Biological Chemistry 258(22): 14025-14033, 1983

Transcription-dependent DNA transactions in the mitochondrial genome of a yeast hypersuppressive petite mutant. Molecular and Cellular Biology 18(5): 2976-2985, 1998

Two modules from the hypersuppressive rho- mitochondrial DNA are required for plasmid replication in yeast. Gene 30(1-3): 47-61, 1984

2 modules from the hypersuppressive rho minus mitochondrial dna are required for plasmid replication in yeast. Gene (Amsterdam) 30(1-3): 47-62, 1984

Genetic, Physical, and Functional Interactions between the Triphosphatase and Guanylyltransferase Components of the Yeast mRna Capping Apparatus. Molecular and Cellular Biology 18(9): 5189-5198, 1998

An essential surface motif (WAQKW) of yeast RNA triphosphatase mediates formation of the mRNA capping enzyme complex with RNA guanylyltransferase. Nucleic Acids Research 27(24): 4671-4678, 1999

Purification and characterization of the mRna capping enzyme Gtp:Rna guanylyltransferase from wheat germ. Biochemistry 21(2): 327-333, 1982

RNA capping by HeLa cell RNA guanylyltransferase. Characterization of a covalent protein-guanylate intermediate. Journal of Biological Chemistry 257(12): 7237-7245, 1982

A conserved domain of yeast RNA triphosphatase flanking the catalytic core regulates self-association and interaction with the guanylyltransferase component of the mRNA capping apparatus. Journal of Biological Chemistry 274(32): 22668-22678, 1999

Mechanism of mRNA capping by vaccinia virus guanylyltransferase: characterization of an enzyme--guanylate intermediate. Proceedings of the National Academy of Sciences of the United States of America 78(1): 187-191, 1981

Purification and characterization of the messenger ribonucleic acid capping enzyme GTP:RNA guanylyltransferase from wheat germ. Biochemistry 21(2): 327-333, 1982

Purification and characterization of the CopB replication control protein, and precise mapping of its target site in the R1 plasmid. Plasmid 15(3): 163-171, 1986

Mutational analysis of yeast RNA triphosphatases highlight a common mechanism of metal-dependent NTP hydrolysis and means of targeting enzymes to pre-mRNAs in vivo by fusion to the guanylyltransferase component of capping apparatus. The Journal of Biological Chemistry 274(41): 865-74, 1999

Mutational analyses of yeast RNA triphosphatases highlight a common mechanism of metal-dependent NTP hydrolysis and a means of targeting enzymes to pre-mRNAs in vivo by fusion to the guanylyltransferase component of the capping apparatus. Journal of Biological Chemistry 274(41): 28865-28874, 1999