Abstract
Escherichia coli rnh mutants defective in RNase H activity display the features of previously described sdrA (stable DNA replication) and dasF (dnaA suppressor) mutants: (i) sustained DNA replication in the absence of protein synthesis, (ii) lack of requirement for dnaA protein and the origin of replication (oriC), and (iii) sensitivity of growth to a rich medium. Both the sdrA mutants (selected for continued DNA replication in the absence of protein synthesis) and the dasF mutants (selected as dnaA suppressors) are defective in RNase H activity, measured in vitro. Furthermore, a 760-base-pair fragment containing the rnh+ structural gene complements the phenotype of each of the rnh, sdrA, and dasF mutants, indicative of a single gene. One function of RNase H in vivo is in the initiation of a cycle of DNA replication at oriC dependent on dnaA+. In keeping with these results, RNase H contributes to the specificity of dnaA protein-dependent replication initiated at oriC in a partially purified enzyme system.
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Selected References
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- Asada K., Sugimoto K., Oka A., Takanami M., Hirota Y. Structure of replication origin of the Escherichia coli K-12 chromosome: the presence of spacer sequences in the ori region carrying information for autonomous replication. Nucleic Acids Res. 1982 Jun 25;10(12):3745–3754. doi: 10.1093/nar/10.12.3745. [DOI] [PMC free article] [PubMed] [Google Scholar]
- BURTON K. A study of the conditions and mechanism of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid. Biochem J. 1956 Feb;62(2):315–323. doi: 10.1042/bj0620315. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bagdasarian M. M., Izakowska M., Bagdasarian M. Suppression of the DnaA phenotype by mutations in the rpoB cistron of ribonucleic acid polymerase in Salmonella typhimurium and Escherichia coli. J Bacteriol. 1977 May;130(2):577–582. doi: 10.1128/jb.130.2.577-582.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bird R. E., Louarn J., Martuscelli J., Caro L. Origin and sequence of chromosome replication in Escherichia coli. J Mol Biol. 1972 Oct 14;70(3):549–566. doi: 10.1016/0022-2836(72)90559-1. [DOI] [PubMed] [Google Scholar]
- Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
- Carl P. L. Escherichia coli mutants with temperature-sensitive synthesis of DNA. Mol Gen Genet. 1970;109(2):107–122. doi: 10.1007/BF00269647. [DOI] [PubMed] [Google Scholar]
- Clewell D. B. Nature of Col E 1 plasmid replication in Escherichia coli in the presence of the chloramphenicol. J Bacteriol. 1972 May;110(2):667–676. doi: 10.1128/jb.110.2.667-676.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dixon N. E., Kornberg A. Protein HU in the enzymatic replication of the chromosomal origin of Escherichia coli. Proc Natl Acad Sci U S A. 1984 Jan;81(2):424–428. doi: 10.1073/pnas.81.2.424. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fuller R. S., Kaguni J. M., Kornberg A. Enzymatic replication of the origin of the Escherichia coli chromosome. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7370–7374. doi: 10.1073/pnas.78.12.7370. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fuller R. S., Kornberg A. Purified dnaA protein in initiation of replication at the Escherichia coli chromosomal origin of replication. Proc Natl Acad Sci U S A. 1983 Oct;80(19):5817–5821. doi: 10.1073/pnas.80.19.5817. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hillenbrand G., Staudenbauer W. L. Discriminatory function of ribonuclease H in the selective initiation of plasmid DNA replication. Nucleic Acids Res. 1982 Feb 11;10(3):833–853. doi: 10.1093/nar/10.3.833. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Itoh T., Tomizawa J. Formation of an RNA primer for initiation of replication of ColE1 DNA by ribonuclease H. Proc Natl Acad Sci U S A. 1980 May;77(5):2450–2454. doi: 10.1073/pnas.77.5.2450. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Itoh T., Tomizawa J. Purification of ribonuclease H as a factor required for initiation of in vitro Co1E1 DNA replication. Nucleic Acids Res. 1982 Oct 11;10(19):5949–5965. doi: 10.1093/nar/10.19.5949. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kaguni J. M., Fuller R. S., Kornberg A. Enzymatic replication of E. coli chromosomal origin is bidirectional. Nature. 1982 Apr 15;296(5858):623–627. doi: 10.1038/296623a0. [DOI] [PubMed] [Google Scholar]
- Kaguni J., LaVerne L. S., Ray D. S. Cloning and expression of the Escherichia coli replication origin in a single-stranded DNA phage. Proc Natl Acad Sci U S A. 1979 Dec;76(12):6250–6254. doi: 10.1073/pnas.76.12.6250. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kanaya S., Crouch R. J. DNA sequence of the gene coding for Escherichia coli ribonuclease H. J Biol Chem. 1983 Jan 25;258(2):1276–1281. [PubMed] [Google Scholar]
- Kogoma T. A novel Escherichia coli mutant capable of DNA replication in the absence of protein synthesis. J Mol Biol. 1978 May 5;121(1):55–69. doi: 10.1016/0022-2836(78)90262-0. [DOI] [PubMed] [Google Scholar]
- Kogoma T., Torrey T. A., Connaughton M. J. Induction of UV-resistant DNA replication in Escherichia coli: induced stable DNA replication as an SOS function. Mol Gen Genet. 1979 Oct 2;176(1):1–9. doi: 10.1007/BF00334288. [DOI] [PubMed] [Google Scholar]
- Kogoma T., von Meyenburg K. The origin of replication, oriC, and the dnaA protein are dispensable in stable DNA replication (sdrA) mutants of Escherichia coli K-12. EMBO J. 1983;2(3):463–468. doi: 10.1002/j.1460-2075.1983.tb01445.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- LARK K. G., REPKO T., HOFFMAN E. J. THE EFFECT OF AMINO ACID DEPRIVATION ON SUBSEQUENT DEOXYRIBONUCLEIC ACID REPLICATION. Biochim Biophys Acta. 1963 Sep 17;76:9–24. [PubMed] [Google Scholar]
- Lark K. G. Evidence for the direct involvement of RNA in the initiation of DNA replication in Escherichia coli 15T. J Mol Biol. 1972 Feb 28;64(1):47–60. doi: 10.1016/0022-2836(72)90320-8. [DOI] [PubMed] [Google Scholar]
- Lark K. G., Lark C. A. recA-dependent DNA replication in the absence of protein synthesis: characteristics of a dominant lethal replication mutation, dnaT, and requirement for recA+ function. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 1):537–549. doi: 10.1101/sqb.1979.043.01.059. [DOI] [PubMed] [Google Scholar]
- Lark K. G., Renger H. Initiation of DNA replication in Escherichia coli 15T-: chronological dissection of three physiological processes required for initiation. J Mol Biol. 1969 Jun 14;42(2):221–235. doi: 10.1016/0022-2836(69)90039-4. [DOI] [PubMed] [Google Scholar]
- MAALOE O., HANAWALT P. C. Thymine deficiency and the normal DNA replication cycle. I. J Mol Biol. 1961 Apr;3:144–155. doi: 10.1016/s0022-2836(61)80041-7. [DOI] [PubMed] [Google Scholar]
- Maki H., Horiuchi T., Sekiguchi M. Structure and expression of the dnaQ mutator and the RNase H genes of Escherichia coli: overlap of the promoter regions. Proc Natl Acad Sci U S A. 1983 Dec;80(23):7137–7141. doi: 10.1073/pnas.80.23.7137. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Masters M., Broda P. Evidence for the bidirectional replications of the Escherichia coli chromosome. Nat New Biol. 1971 Aug 4;232(31):137–140. doi: 10.1038/newbio232137a0. [DOI] [PubMed] [Google Scholar]
- Meijer M., Beck E., Hansen F. G., Bergmans H. E., Messer W., von Meyenburg K., Schaller H. Nucleotide sequence of the origin of replication of the Escherichia coli K-12 chromosome. Proc Natl Acad Sci U S A. 1979 Feb;76(2):580–584. doi: 10.1073/pnas.76.2.580. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Naito S., Kitani T., Ogawa T., Okazaki T., Uchida H. Escherichia coli mutants suppressing replication-defective mutations of the ColE1 plasmid. Proc Natl Acad Sci U S A. 1984 Jan;81(2):550–554. doi: 10.1073/pnas.81.2.550. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Oka A., Sugimoto K., Takanami M., Hirota Y. Replication origin of the Escherichia coli K-12 chromosome: the size and structure of the minimum DNA segment carrying the information for autonomous replication. Mol Gen Genet. 1980 Apr;178(1):9–20. doi: 10.1007/BF00267207. [DOI] [PubMed] [Google Scholar]
- Prescott D. M., Kuempel P. L. Bidirectional replication of the chromosome in Escherichia coli. Proc Natl Acad Sci U S A. 1972 Oct;69(10):2842–2845. doi: 10.1073/pnas.69.10.2842. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sugimoto K., Oka A., Sugisaki H., Takanami M., Nishimura A., Yasuda Y., Hirota Y. Nucleotide sequence of Escherichia coli K-12 replication origin. Proc Natl Acad Sci U S A. 1979 Feb;76(2):575–579. doi: 10.1073/pnas.76.2.575. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Torrey T. A., Kogoma T. Suppressor mutations (rin) that specifically suppress the recA+ dependence of stable DNA replication in Escherichia coliK-12. Mol Gen Genet. 1982;187(2):225–230. doi: 10.1007/BF00331121. [DOI] [PubMed] [Google Scholar]
- Vicuna R., Hurwitz J., Wallace S., Girard M. Selective inhibition of in vitro DNA synthesis dependent on phiX174 compared with fd DNA. I. Protein requirements for selective inhibition. J Biol Chem. 1977 Apr 25;252(8):2524–2533. [PubMed] [Google Scholar]
- Zyskind J. W., Smith D. W. NOVEL Escherichia coli dnaB mutant: direct involvement of the dnaB252 gene product in the synthesis of an origin-ribonucleic acid species during initiaion of a round of deoxyribonucleic acid replication. J Bacteriol. 1977 Mar;129(3):1476–1486. doi: 10.1128/jb.129.3.1476-1486.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]