Abstract
phiX174 DNA-dependent DNA synthesis is catalyzed in vitro by the combination of at least 11 purified protein fractions: dnaB, dnaC(D), and dnaG gene products, DNA polymerase III, DNA elongation factors I and II, DNA binding protein, and replication factors W, X, Y, and Z. The reaction requires ATP, 4 dNTPs, and Mg+2 and is specific for phiX174 (or phiXahb) DNA. Purified replication factor Y contains phiX174 (or phiXahb) DNA-dependent ATPase (or dATPase) activity. The ATPase activity is poorly stimulated by other single-stranded DNA, by double-stranded DNA, or by RNA. The products of the phiX174 DNA-dependent ATPase activity of factor Y are Pi and ADP (or dADP). The association of phiX174 DNA-dependent ATPase activity with factor Y was shown in the following ways: (a) the two activities copurified with a constant ratio; (b) they comigrated on native polyacrylamide gel electrophoresis; (c) both activities were heat-inactivated at the same rate; and (d) both showed identical patterns of N-ethylmaleimide sensitivity.
Full text
PDF![3342](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f12/432988/fd1130aa49fe/pnas00052-0084.png)
![3343](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f12/432988/297bf15a46d5/pnas00052-0085.png)
![3344](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f12/432988/c370585b4514/pnas00052-0086.png)
![3345](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f12/432988/8bfcfbbb192e/pnas00052-0087.png)
![3346](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f12/432988/c12698541829/pnas00052-0088.png)
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bone D. R., Dowel C. E. A mutant of bacteriophage øX174 which infects E. coli K12 strains. I. Isolation and partial characterization of øXtB. Virology. 1973 Apr;52(2):319–329. [PubMed] [Google Scholar]
- Bradley D. E. A comparative study of some properties of the phi-X174 type bacteriophages. Can J Microbiol. 1970 Oct;16(10):965–971. doi: 10.1139/m70-165. [DOI] [PubMed] [Google Scholar]
- Buttin G., Wright M. Enzymatic DNA degradation in E. coli: its relationship to synthetic processes at the chromosome level. Cold Spring Harb Symp Quant Biol. 1968;33:259–269. doi: 10.1101/sqb.1968.033.01.030. [DOI] [PubMed] [Google Scholar]
- CONWAY T. W., LIPMANN F. CHARACTERIZATION OF A RIBOSOME-LINKED GUANOSINE TRIPHOSPHATASE IN ESCHERICHIA COLI EXTRACTS. Proc Natl Acad Sci U S A. 1964 Dec;52:1462–1469. doi: 10.1073/pnas.52.6.1462. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ebisuzaki K., Behme M. T., Senior C., Shannon D., Dunn D. An alternative approach to the study of new enzymatic reactions involving DNA (DNA-dependent ATPases-purification-properties-E. coli). Proc Natl Acad Sci U S A. 1972 Feb;69(2):515–519. doi: 10.1073/pnas.69.2.515. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gefter M. L., Hirota Y., Kornberg T., Wechsler J. A., Barnoux C. Analysis of DNA polymerases II and 3 in mutants of Escherichia coli thermosensitive for DNA synthesis. Proc Natl Acad Sci U S A. 1971 Dec;68(12):3150–3153. doi: 10.1073/pnas.68.12.3150. [DOI] [PMC free article] [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]
- Godson G. N. Evolution of phi-chi 174. Isolation of four new phi-chi-like phages and comparison with phi-chi 174. Virology. 1974 Mar;58(1):272–289. doi: 10.1016/0042-6822(74)90161-5. [DOI] [PubMed] [Google Scholar]
- Hurwitz J., Wickner S. Involvement of two protein factors and ATP in in vitro DNA synthesis catalyzed by DNA polymerase 3 of Escherichia coli. Proc Natl Acad Sci U S A. 1974 Jan;71(1):6–10. doi: 10.1073/pnas.71.1.6. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hurwitz J., Wickner S., Wright M. Studies on in vitro DNA synthesis. II. Isolation of a protein which stimulates deoxynucleotide incorporation catalyzed by DNA polymerase of E. coli. Biochem Biophys Res Commun. 1973 Mar 17;51(2):257–267. doi: 10.1016/0006-291x(73)91251-5. [DOI] [PubMed] [Google Scholar]
- JOVIN T., CHRAMBACH A., NAUGHTON M. A. AN APPARATUS FOR PREPARATIVE TEMPERATURE-REGULATED POLYACRYLAMIDE GEL ELECTROPHORESIS. Anal Biochem. 1964 Nov;9:351–369. doi: 10.1016/0003-2697(64)90192-7. [DOI] [PubMed] [Google Scholar]
- Livingston D. M., Hinkle D. C., Richardson C. C. Deoxyribonucleic acid polymerase III of Escherichia coli. Purification and properties. J Biol Chem. 1975 Jan 25;250(2):461–469. [PubMed] [Google Scholar]
- Molineux I. J., Friedman S., Gefter M. L. Purification and properties of the Escherichia coli deoxyribonucleic acid-unwinding protein. Effects on deoxyribonucleic acid synthesis in vitro. J Biol Chem. 1974 Oct 10;249(19):6090–6098. [PubMed] [Google Scholar]
- Nüsslein V., Otto B., Bonhoeffer F., Schaller H. Function of DNA polymerase 3 in DNA replication. Nat New Biol. 1971 Dec 29;234(52):285–286. doi: 10.1038/newbio234285a0. [DOI] [PubMed] [Google Scholar]
- Schekman R., Weiner A., Kornberg A. Multienzyme systems of DNA replication. Science. 1974 Dec 13;186(4168):987–993. doi: 10.1126/science.186.4168.987. [DOI] [PubMed] [Google Scholar]
- Sigal N., Delius H., Kornberg T., Gefter M. L., Alberts B. A DNA-unwinding protein isolated from Escherichia coli: its interaction with DNA and with DNA polymerases. Proc Natl Acad Sci U S A. 1972 Dec;69(12):3537–3541. doi: 10.1073/pnas.69.12.3537. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vito C. C., Primrose S. B., Dowell C. E. Growth of a capsid mutant of bacteriophage phi X174 in a temperature-sensitive strain of Escherichia coli. J Virol. 1975 Feb;15(2):281–287. doi: 10.1128/jvi.15.2.281-287.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Weiner J. H., Bertsch L. L., Kornberg A. The deoxyribonucleic acid unwinding protein of Escherichia coli. Properties and functions in replication. J Biol Chem. 1975 Mar 25;250(6):1972–1980. [PubMed] [Google Scholar]
- Wickner S., Berkower I., Wright M., Hurwitz J. Studies on in vitro DNA synthesis: purification of dna C gene product containing dna D activity from Escherichia coli. Proc Natl Acad Sci U S A. 1973 Aug;70(8):2369–2373. doi: 10.1073/pnas.70.8.2369. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wickner S., Hurwitz J. Conversion of phiX174 viral DNA to double-stranded form by purified Escherichia coli proteins. Proc Natl Acad Sci U S A. 1974 Oct;71(10):4120–4124. doi: 10.1073/pnas.71.10.4120. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wickner S., Hurwitz J. Interaction of Escherichia coli dnaB and dnaC(D) gene products in vitro. Proc Natl Acad Sci U S A. 1975 Mar;72(3):921–925. doi: 10.1073/pnas.72.3.921. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wickner S., Wright M., Hurwitz J. Association of DNA-dependent and -independent ribonucleoside triphosphatase activities with dnaB gene product of Escherichia coli. Proc Natl Acad Sci U S A. 1974 Mar;71(3):783–787. doi: 10.1073/pnas.71.3.783. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wickner S., Wright M., Hurwitz J. Studies on in vitro DNA synthesis. Purification of the dna G gene product from Escherichia coli. Proc Natl Acad Sci U S A. 1973 May;70(5):1613–1618. doi: 10.1073/pnas.70.5.1613. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wright M., Wickner S., Hurwitz J. Studies on in vitro DNA synthesis. Isolation of DNA B gene product from Escherichia coli. Proc Natl Acad Sci U S A. 1973 Nov;70(11):3120–3124. doi: 10.1073/pnas.70.11.3120. [DOI] [PMC free article] [PubMed] [Google Scholar]