Targeting a truncated Ho-endonuclease of yeast to novel DNA sites with foreign zinc fingers

Edna Nahon, Dina Raveh

Research output: Contribution to journalArticlepeer-review

21 Scopus citations


Ho-endonuclease of the yeast, Saccharomyces cerevisiae, initiates a mating type switch by making a site-specific double strand break in the mating type gene, MAT. Ho is a dodecamer endonuclease and shares six of the seven intein motifs with PI-Scel endonuclease, an intein encoded by the VMAI gene. We show that a 113 residue truncated Ho-endonuclease starting at intein motif C initiates a mating type switch in yeast. Ho is the only dodecamer endonuclease with zinc fingers. To see whether they have a role in determining site specificity we exchanged them for zinc fingers of the yeast transcription factor, Swi5. A chimeric endonuclease comprising the dodecamer motifs of Ho (C-E) and the zinc finger domain of Swi5 cleaves a Swi5 substrate plasmid in vivo. A similar chimera with the zinc fingers of Spl cleaves a GC box rich substrate plasmid. These experiments delineate a catalytic fragment of Ho-endonuclease that can be fused to various DNA binding moieties in the design of chimeric endonucleases with new site specificities.

Original languageEnglish
Pages (from-to)1233-1239
Number of pages7
JournalNucleic Acids Research
Issue number5
StatePublished - 1 Mar 1998

ASJC Scopus subject areas

  • Genetics


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