Resolution of holliday junction analogs by T4 endonuclease VII can be directed by substrate structure

John E. Mueller, Colin J. Newton, Frank Jensch, Börries Kemper, Richard P. Cunningham, Neville R. Kallenbach, Nadrian Seeman

Research output: Contribution to journalArticle

Abstract

Endonuclease VII is an enzyme from bacteriophage T4 capable of resolving four-arm Holliday junction intermediates in recombination. Since natural Holliday junctions have homologous (2-fold) sequence symmetry, they can branch migrate, creating a population of substrates that have the branch point at different sites. We have explored the substrate requirements of endonuclease VII by using immobile analogs of Holliday junctions that lack this homology, thereby situating the branch point at a fixed site in the molecule. We have found that immobile junctions whose double-helical arms contain fewer than nine nucleotide pairs do not serve as substrates for resolution by endonuclease VII. Scission of substrates with 2-fold symmetrically elongated arms produces resolution products that are a function of the particular arms that are lengthened. We have confirmed that the scission products are those of resolution, rather than nicking of individual strands, by using shamrock junction molecules formed from a single oligonucleotide strand. A combination of end-labeled and internally labeled shamrock molecules has been used to demonstrate that all of the scission is due to coordinated cleavage of DNA on opposite sides of the junction, 3′ to the branch point. Endonuclease VII is known to cleave the crossover strands of Holliday junctions in this fashion. The relationship of the long arms to the cleavage direction suggests that the portion of the enzyme which requires the minimum arm length interacts with the pair of arms containing the 3′ portion of the crossover strands on the bound surface of the antiparallel junction.

Original languageEnglish (US)
Pages (from-to)13918-13924
Number of pages7
JournalJournal of Biological Chemistry
Volume265
Issue number23
StatePublished - Aug 15 1990

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Cruciform DNA
Substrates
Molecules
Bacteriophage T4
DNA Cleavage
Bacteriophages
Enzymes
Oligonucleotides
Genetic Recombination
Nucleotides
endodeoxyribonuclease VII
DNA
Population

ASJC Scopus subject areas

  • Biochemistry

Cite this

Mueller, J. E., Newton, C. J., Jensch, F., Kemper, B., Cunningham, R. P., Kallenbach, N. R., & Seeman, N. (1990). Resolution of holliday junction analogs by T4 endonuclease VII can be directed by substrate structure. Journal of Biological Chemistry, 265(23), 13918-13924.

Resolution of holliday junction analogs by T4 endonuclease VII can be directed by substrate structure. / Mueller, John E.; Newton, Colin J.; Jensch, Frank; Kemper, Börries; Cunningham, Richard P.; Kallenbach, Neville R.; Seeman, Nadrian.

In: Journal of Biological Chemistry, Vol. 265, No. 23, 15.08.1990, p. 13918-13924.

Research output: Contribution to journalArticle

Mueller, JE, Newton, CJ, Jensch, F, Kemper, B, Cunningham, RP, Kallenbach, NR & Seeman, N 1990, 'Resolution of holliday junction analogs by T4 endonuclease VII can be directed by substrate structure', Journal of Biological Chemistry, vol. 265, no. 23, pp. 13918-13924.
Mueller JE, Newton CJ, Jensch F, Kemper B, Cunningham RP, Kallenbach NR et al. Resolution of holliday junction analogs by T4 endonuclease VII can be directed by substrate structure. Journal of Biological Chemistry. 1990 Aug 15;265(23):13918-13924.
Mueller, John E. ; Newton, Colin J. ; Jensch, Frank ; Kemper, Börries ; Cunningham, Richard P. ; Kallenbach, Neville R. ; Seeman, Nadrian. / Resolution of holliday junction analogs by T4 endonuclease VII can be directed by substrate structure. In: Journal of Biological Chemistry. 1990 ; Vol. 265, No. 23. pp. 13918-13924.
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