Abstract
We have analyzed and compared the responses of the three major HeLa cell DNA polymerases (alpha, beta, and gamma) to a HeLa DNA template with short RNA or DNA primers hybridized to it. Only DNA polymerase alpha is able to synthesize DNA covalently bonded to the RNA primer via a 3' yields 5' phosphodiester bond. 32P transfer experiments showed that all combinations of ribo- and deoxyribonucleotides are represented in the RNA-DNA linkages but their distribution is nonrandom. The RNA-DNA linked molecules base-paired to a HeLa DNA template strand represent a possible "natural" in vitro primer-template for DNA polymerases and can be extended by all three DNA polymerases (alpha, beta, and gamma). These findings indicate that DNA polymerases beta and gamma are capable of DNA-primed but not RNA-PRIMED DNA synthesis, while DNA polymerase alpha is capable of both RNA-primed and DAN-primed DNA synthesis.
Full text
PDFSelected References
These references are in PubMed. This may not be the complete list of references from this article.
- Chang L. M., Bollum F. J. A chemical model for transcriptional initiation of DNA replication. Biochem Biophys Res Commun. 1972 Feb 16;46(3):1354–1360. doi: 10.1016/s0006-291x(72)80124-4. [DOI] [PubMed] [Google Scholar]
- Chang L. M., Bollum F. J. Low molecular weight deoxyribonucleic acid polymerase in mammalian cells. J Biol Chem. 1971 Sep 25;246(18):5835–5837. [PubMed] [Google Scholar]
- Eliasson R., Martin R., Reichard P. Characterization of the RNA initiating the discontinuous synthesis of polyoma DNA. Biochem Biophys Res Commun. 1974 Jul 10;59(1):307–313. doi: 10.1016/s0006-291x(74)80207-x. [DOI] [PubMed] [Google Scholar]
- Fansler B. S. Eukaryotic DNA polymerases: their association with the nucleus and relationship to DNA replication. Int Rev Cytol. 1974;Suppl 4:363–415. [PubMed] [Google Scholar]
- Fridlender B., Fry M., Bolden A., Weissbach A. A new synthetic RNA-dependent DNA polymerase from human tissue culture cells (HeLa-fibroblast-synthetic oligonucleotides-template-purified enzymes). Proc Natl Acad Sci U S A. 1972 Feb;69(2):452–455. doi: 10.1073/pnas.69.2.452. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fry M., Weissbach A. A new deoxyribonucleic acid dependent deoxyribonucleic acid polymerase from HeLa cell mitochondria. Biochemistry. 1973 Sep 11;12(19):3602–3608. doi: 10.1021/bi00743a005. [DOI] [PubMed] [Google Scholar]
- Geider K., Kornberg A. Conversion of the M13 viral single strand to the double-stranded replicative forms by purified proteins. J Biol Chem. 1974 Jul 10;249(13):3999–4005. [PubMed] [Google Scholar]
- Hirose S., Okazaki R., Tamanoi F. Mechanism of DNA chain growth. XI. Structure of RNA-linked DNA fragments of Escherichia coli. J Mol Biol. 1973 Jul 15;77(4):501–517. doi: 10.1016/0022-2836(73)90219-2. [DOI] [PubMed] [Google Scholar]
- Keller W. RNA-primed DNA synthesis in vitro. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1560–1564. doi: 10.1073/pnas.69.6.1560. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Magnusson G., Pigiet V., Winnacker E. L., Abrams R., Reichard P. RNA-linked short DNA fragments during polyoma replication. Proc Natl Acad Sci U S A. 1973 Feb;70(2):412–415. doi: 10.1073/pnas.70.2.412. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Maïa J. C.C., Rougeon F., Chapeville F. Chick embryo poly (rA:dT)-dependent DNA polymerase. FEBS Lett. 1971 Oct 15;18(1):130–134. doi: 10.1016/0014-5793(71)80427-1. [DOI] [PubMed] [Google Scholar]
- Pigiet V., Eliasson R., Reichard P. Replication of polyoma DNA in isolated nuclei. 3. The nucleotide sequence at the RNA-DNA junction of nascent strands. J Mol Biol. 1974 Mar 25;84(1):197–216. doi: 10.1016/0022-2836(74)90222-8. [DOI] [PubMed] [Google Scholar]
- Schlabach A., Fridlender B., Bolden A., Weissbach A. DNA-dependent DNA polymerases from HeLa cell nuclei. II. Template and substrate utilization. Biochem Biophys Res Commun. 1971 Aug 20;44(4):879–885. doi: 10.1016/0006-291x(71)90793-5. [DOI] [PubMed] [Google Scholar]
- Spadari S., Weissbach A. HeLa cell R-deoxyribonucleic acid polymerases. Separation and characterization of two enzymatic activities. J Biol Chem. 1974 Sep 25;249(18):5809–5815. [PubMed] [Google Scholar]
- Sugino A., Okazaki R. RNA-linked DNA fragments in vitro. Proc Natl Acad Sci U S A. 1973 Jan;70(1):88–92. doi: 10.1073/pnas.70.1.88. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Weissbach A., Schlabach A., Fridlender B., Bolden A. DNA polymerases from human cells. Nat New Biol. 1971 Jun 9;231(23):167–170. doi: 10.1038/newbio231167a0. [DOI] [PubMed] [Google Scholar]
- YONEDA M., BOLLUM F. J. DEOXYNUCLEOTIDE-POLYMERIZING ENZYMES OF CALF THYMUS GLAND. I. LARGE SCALE PURIFICATION OF TERMINAL AND REPLICATIVE DEOXYNUCLEOTIDYL TRANSFERASES. J Biol Chem. 1965 Aug;240:3385–3391. [PubMed] [Google Scholar]