Bacteriophage T4 encodes an RNase H which removes RNA primers made by the T4 DNA replication system in vitro
- PMID: 1703156
Bacteriophage T4 encodes an RNase H which removes RNA primers made by the T4 DNA replication system in vitro
Abstract
RNase H activity increases markedly after bacteriophage T4 infection of Escherichia coli MIC2003, an RNase H-deficient host. We have extensively purified the RNase H from these T4-infected cells and have shown that the RNase H activity copurifies with a 5' to 3' DNA exonuclease activity. The N-terminal sequence of a 35-kDa protein copurifying with the RNase H activity matches the terminus of the predicted product of an open reading frame (designated ORF A or 33.2) upstream of T4 gene 33, identified previously by Hahn and co-workers (Hahn, S., Kruse, U., and Rüger, W. (1986) Nucleic Acids Res. 14, 9311-9327). Plasmids containing ORF A under the control of the T7 promoter express RNase H and 5' to 3' DNA exonuclease activities as well as a protein that comigrates on sodium dodecyl sulfate-polyacrylamide gels with the 35-kDa protein present in the RNase H purified from T4-infected cells. T4 RNase H removes the pentamer RNA primers from DNA chains initiated by the T4 primase-helicase (gene products 61 and 41). Addition of T4 RNase H and T4 DNA ligase leads to extensive joining of discontinuous lagging strand fragments in the T4 DNA replication system in vitro.
Similar articles
-
Trypsin cleavage in the COOH terminus of the bacteriophage T4 gene 41 DNA helicase alters the primase-helicase activities of the T4 replication complex in vitro.J Biol Chem. 1989 Mar 15;264(8):4732-9. J Biol Chem. 1989. PMID: 2466840
-
Effects of the bacteriophage T4 gene 41 and gene 32 proteins on RNA primer synthesis: coupling of leading- and lagging-strand DNA synthesis at a replication fork.Biochemistry. 1990 Feb 20;29(7):1791-8. doi: 10.1021/bi00459a018. Biochemistry. 1990. PMID: 2158814
-
Analysis of five presumptive protein-coding sequences clustered between the primosome genes, 41 and 61, of bacteriophages T4, T2, and T6.J Virol. 1993 Apr;67(4):2305-16. doi: 10.1128/JVI.67.4.2305-2316.1993. J Virol. 1993. PMID: 8383243 Free PMC article.
-
Either bacteriophage T4 RNase H or Escherichia coli DNA polymerase I is essential for phage replication.J Bacteriol. 1996 Dec;178(23):6772-7. doi: 10.1128/jb.178.23.6772-6777.1996. J Bacteriol. 1996. PMID: 8955295 Free PMC article.
-
Protein-protein interactions at a DNA replication fork: bacteriophage T4 as a model.FASEB J. 1992 Feb 1;6(3):871-8. doi: 10.1096/fasebj.6.3.1310946. FASEB J. 1992. PMID: 1310946 Review.
Cited by
-
Structural conservation of the PIN domain active site across all domains of life.Protein Sci. 2017 Aug;26(8):1474-1492. doi: 10.1002/pro.3193. Epub 2017 May 24. Protein Sci. 2017. PMID: 28508407 Free PMC article. Review.
-
Saccharomyces cerevisiae RNase H(35) functions in RNA primer removal during lagging-strand DNA synthesis, most efficiently in cooperation with Rad27 nuclease.Mol Cell Biol. 1999 Dec;19(12):8361-71. doi: 10.1128/MCB.19.12.8361. Mol Cell Biol. 1999. PMID: 10567561 Free PMC article.
-
Characterization of DNA synthesis catalyzed by bacteriophage T4 replication complexes reconstituted on synthetic circular substrates.Nucleic Acids Res. 2002 Oct 15;30(20):4387-97. doi: 10.1093/nar/gkf576. Nucleic Acids Res. 2002. PMID: 12384585 Free PMC article.
-
Structural analysis of bacteriophage T4 DNA replication: a review in the Virology Journal series on bacteriophage T4 and its relatives.Virol J. 2010 Dec 3;7:359. doi: 10.1186/1743-422X-7-359. Virol J. 2010. PMID: 21129204 Free PMC article. Review.
-
Molecular cloning of a ribonuclease H (RNase HI) gene from an extreme thermophile Thermus thermophilus HB8: a thermostable RNase H can functionally replace the Escherichia coli enzyme in vivo.Nucleic Acids Res. 1991 Aug 25;19(16):4443-9. doi: 10.1093/nar/19.16.4443. Nucleic Acids Res. 1991. PMID: 1653414 Free PMC article.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Molecular Biology Databases
Research Materials
