HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Brennan, C. A. Articles by Gumport, R. I. Search for Related Content PubMed PubMed Citation Articles by Brennan, C. A. Articles by Gumport, R. I. Social Bookmarking                      What's this?

J. Biol. Chem., Vol. 261, Issue 16, 7270-7278, 06, 1986

The effects of base analogue substitutions on the cleavage by the EcoRI restriction endonuclease of octadeoxyribonucleotides containing modified EcoRI recognition sequences

CA Brennan, MD Van Cleve and RI Gumport

It has been proposed that recognition of specific DNA sequences by proteins is accomplished by hydrogen bond formation between the protein and particular groups that are accessible in the major and minor grooves of the DNA. We have examined the DNA-protein interactions involved in the recognition of the hexameric DNA sequence, GAATTC, by the EcoRI restriction endonuclease by using derivatives of an oligodeoxyribonucleotide that contain a variety of base analogues. The base analogues hypoxanthine, 2-aminopurine, 2,6-diaminopurine, N6- methyladenine, 5-bromouracil, uracil, 5-bromocytosine, and 5- methylcytosine were incorporated as single substitutions into the octadeoxyribonucleotide d(pG-G-A-A-T-T-C-C). The effects of the substitutions on the interactions between the EcoRI endonuclease and its recognition sequence were monitored by determining the steady state kinetic values of the hydrolysis reaction. The substitutions resulted in effects that varied from complete inactivity to enhanced reactivity. The enzyme exhibited Michaelis-Menten kinetics with those substrates that were reactive, whereas octanucleotide analogues containing N6- methyladenine at either adenine position, uracil at the second thymine position, or 5-bromocytosine or 5-methylcytosine at the cytosine position were unreactive. The results are discussed in terms of possible effects on interactions between the enzyme and its recognition site during the reaction. An accompanying paper presents the results of a similar study using these oligonucleotides with the EcoRI modification methylase.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				V. Valinluck and L. C. Sowers Inflammation-Mediated Cytosine Damage: A Mechanistic Link between Inflammation and the Epigenetic Alterations in Human Cancers Cancer Res., June 15, 2007; 67(12): 5583 - 5586. [Abstract] [Full Text] [PDF]

				B. I. Kankia A real-time assay for monitoring nucleic acid cleavage by quadruplex formation Nucleic Acids Res., November 6, 2006; 34(20): e141 - e141. [Abstract] [Full Text] [PDF]

				V. Valinluck, P. Liu, J. I. Kang Jr, A. Burdzy, and L. C. Sowers 5-Halogenated pyrimidine lesions within a CpG sequence context mimic 5-methylcytosine by enhancing the binding of the methyl-CpG-binding domain of methyl-CpG-binding protein 2 (MeCP2) Nucleic Acids Res., May 25, 2005; 33(9): 3057 - 3064. [Abstract] [Full Text] [PDF]

				V. Valinluck, H.-H. Tsai, D. K. Rogstad, A. Burdzy, A. Bird, and L. C. Sowers Oxidative damage to methyl-CpG sequences inhibits the binding of the methyl-CpG binding domain (MBD) of methyl-CpG binding protein 2 (MeCP2) Nucleic Acids Res., August 9, 2004; 32(14): 4100 - 4108. [Abstract] [Full Text] [PDF]

				S. Chandrashekaran, U. H. Manjunatha, and V. Nagaraja KpnI restriction endonuclease and methyltransferase exhibit contrasting mode of sequence recognition Nucleic Acids Res., June 10, 2004; 32(10): 3148 - 3155. [Abstract] [Full Text] [PDF]

				A. MACHWE, D. K. ORREN, and V. A. BOHR Accelerated methylation of ribosomal RNA genes during the cellular senescence of Werner syndrome fibroblasts FASEB J, September 1, 2000; 14(12): 1715 - 1724. [Abstract] [Full Text]

				U. Kettling, A. Koltermann, P. Schwille, and M. Eigen Real-time enzyme kinetics monitored by dual-color fluorescence cross-correlation spectroscopy PNAS, February 17, 1998; 95(4): 1416 - 1420. [Abstract] [Full Text] [PDF]

				B. W. Allan, J. M. Beechem, W. M. Lindstrom, and N. O. Reich Direct Real Time Observation of Base Flipping by the EcoRI DNA Methyltransferase J. Biol. Chem., January 23, 1998; 273(4): 2368 - 2373. [Abstract] [Full Text] [PDF]

				L. J. Duggan, T. M. Hill, S. Wu, K. Garrison, X. Zhang, and P. A. Gottlieb Using Modified Nucleotides to Map the DNA Determinants of the Tus-TerB Complex, the Protein-DNA Interaction Associated with Termination of Replication in Escherichia coli J. Biol. Chem., November 24, 1995; 270(47): 28049 - 28054. [Abstract] [Full Text] [PDF]

				B. E. Withers and J. C. Dunbar Sequence-specific DNA Recognition by the SmaI Endonuclease J. Biol. Chem., March 24, 1995; 270(12): 6496 - 6504. [Abstract] [Full Text] [PDF]

				D. Lesser, M. Kurpiewski, and L Jen-Jacobson The energetic basis of specificity in the Eco RI endonuclease--DNA interaction Science, November 9, 1990; 250(4982): 776 - 786. [Abstract] [PDF]