Search tips
Search criteria 


Logo of narLink to Publisher's site
Nucleic Acids Res. 1988 May 11; 16(9): 3997–4008.
PMCID: PMC336570

EcoRII can be activated to cleave refractory DNA recognition sites.


EcoRII restriction sites [5'-CC(A/T)GG] in phage T3 and T7 DNA are refractory to cleavage by EcoRII, but become sensitive to cleavage in the presence of DNAs which contain an abundance of EcoRII sensitive sites (e.g. pBR322 or lambda DNA). Studies using fragments of pBR322 containing different numbers of EcoRII sites show that the susceptibility to EcoRII cleavage is proportional to the number of sites in the individual fragment. We postulate that EcoRII is the prototype of restriction endonucleases which require at least 2 simultaneously bound substrate sites for their activation. EcoRII sites are refractory when they occur at relatively low frequency in the DNA. The restriction enzyme can be activated by DNA with a higher frequency of sites.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.6M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Krüger DH, Bickle TA. Bacteriophage survival: multiple mechanisms for avoiding the deoxyribonucleic acid restriction systems of their hosts. Microbiol Rev. 1983 Sep;47(3):345–360. [PMC free article] [PubMed]
  • Brennan CA, Van Cleve MD, Gumport RI. The effects of base analogue substitutions on the cleavage by the EcoRI restriction endonuclease of octadeoxyribonucleotides containing modified EcoRI recognition sequences. J Biol Chem. 1986 Jun 5;261(16):7270–7278. [PubMed]
  • Yolov AA, Vinogradova MN, Gromova ES, Rosenthal A, Cech D, Veiko VP, Metelev VG, Kosykh VG, Buryanov YI, Bayev AA, et al. Interaction of EcoRII restriction and modification enzymes with synthetic DNA fragments. VI. The binding and cleavage of substrates containing nucleotide analogs. Nucleic Acids Res. 1985 Dec 20;13(24):8983–8998. [PMC free article] [PubMed]
  • Bron S, Luxen E, Venema G. Resistance of bacteriophage H1 to restriction and modification by Bacillus subtilis R. J Virol. 1983 Jun;46(3):703–708. [PMC free article] [PubMed]
  • Busslinger M, deBoer E, Wright S, Grosveld FG, Flavell RA. The sequence GGCmCGG is resistant to MspI cleavage. Nucleic Acids Res. 1983 Jun 11;11(11):3559–3569. [PMC free article] [PubMed]
  • Thomas M, Davis RW. Studies on the cleavage of bacteriophage lambda DNA with EcoRI Restriction endonuclease. J Mol Biol. 1975 Jan 25;91(3):315–328. [PubMed]
  • Krüger DH, Schroeder C, Reuter M, Bogdarina IG, Buryanov YI, Bickle TA. DNA methylation of bacterial viruses T3 and T7 by different DNA methylases in Escherichia coli K12 cells. Eur J Biochem. 1985 Jul 15;150(2):323–330. [PubMed]
  • Nelson M, McClelland M. The effect of site-specific methylation on restriction-modification enzymes. Nucleic Acids Res. 1987;15 (Suppl):r219–r230. [PMC free article] [PubMed]
  • Vovis GF, Horiuchi K, Zinder ND. Endonuclease R-EcoRII restriction of bacteriophage f1 DNA in vitro: ordering of genes V and VII, location of an RNA promotor for gene VIII. J Virol. 1975 Sep;16(3):674–684. [PMC free article] [PubMed]
  • Hattman S, Gribbin C, Hutchison CA., 3rd In vivo methylation of bacteriophage phi X174 DNA. J Virol. 1979 Dec;32(3):845–851. [PMC free article] [PubMed]
  • O'Farrell PH, Kutter E, Nakanishi M. A restriction map of the bacteriophage T4 genome. Mol Gen Genet. 1980;179(2):421–435. [PubMed]
  • Sharp PA, Sugden B, Sambrook J. Detection of two restriction endonuclease activities in Haemophilus parainfluenzae using analytical agarose--ethidium bromide electrophoresis. Biochemistry. 1973 Jul 31;12(16):3055–3063. [PubMed]
  • Dretzen G, Bellard M, Sassone-Corsi P, Chambon P. A reliable method for the recovery of DNA fragments from agarose and acrylamide gels. Anal Biochem. 1981 Apr;112(2):295–298. [PubMed]
  • Pingoud A, Urbanke C, Alves J, Ehbrecht HJ, Zabeau M, Gualerzi C. Effect of polyamines and basic proteins on cleavage of DNA by restriction endonucleases. Biochemistry. 1984 Nov 20;23(24):5697–5703. [PubMed]
  • Sutcliffe JG. Complete nucleotide sequence of the Escherichia coli plasmid pBR322. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 1):77–90. [PubMed]
  • Peden KW. Revised sequence of the tetracycline-resistance gene of pBR322. Gene. 1983 May-Jun;22(2-3):277–280. [PubMed]
  • Hughes SG. The sensitivity of bacteriophage lambda DNA to restriction endonuclease RII. J Mol Biol. 1975 Nov 5;98(3):645–647. [PubMed]
  • Guseinov OA, Bogdarina IG, Kosykh VG, Bur'ianov Ia I, Baev AA. Chuvstvitel'nost' khromosomnykh i plazmidnykh DNK shtammov E. coli k restriktsionnoi endonykleaze Eco RII. Biokhimiia. 1978 Sep;43(9):1718–1720. [PubMed]
  • Vinogradova MN, Gromova ES, Purmal' AA, Kosykh VG, Shabarova ZA. Vzaimodeistvie fermentov restriktsii i mofifikatsii EcoRII s sinteticheskimi fragmentami DNK. VII. Izuchenie kompleksoobrazovaniia éndonukleazy EcoRII s substratami, soderzhashchimi prirodnye i modifitsirovannye uchastki uznavaniia. Mol Biol (Mosk) 1986 Sep-Oct;20(5):1329–1336. [PubMed]
  • Dunn JJ, Studier FW. Complete nucleotide sequence of bacteriophage T7 DNA and the locations of T7 genetic elements. J Mol Biol. 1983 Jun 5;166(4):477–535. [PubMed]
  • Yamada M, Fujisawa H, Kato H, Hamada K, Minagawa T. Cloning and sequencing of the genetic right end of bacteriophage T3 DNA. Virology. 1986 Jun;151(2):350–361. [PubMed]
  • Schroeder C, Jurkschat H, Meisel A, Reich JG, Krüger D. Unusual occurrence of EcoP1 and EcoP15 recognition sites and counterselection of type II methylation and restriction sequences in bacteriophage T7 DNA. Gene. 1986;45(1):77–86. [PubMed]
  • Piekarowicz A. Preferential cleavage by restriction endonuclease HinfIII. Acta Biochim Pol. 1984;31(4):453–464. [PubMed]
  • Yanisch-Perron C, Vieira J, Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. [PubMed]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press