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GENES CELLS (2003) 8, 587-602.
Copyright © 2003 Blackwell Publishing or its licensors

Original Article

VDE-initiated intein homing in Saccharomyces cerevisiae proceeds in a meiotic recombination-like manner

T Fukuda, S Nogami, and Y Ohya

BACKGROUND: Inteins and group I introns found in prokaryotic and eukaryotic organisms occasionally behave as mobile genetic elements. During meiosis of the yeast Saccharomyces cerevisiae, the site-specific endonuclease encoded by VMA1 intein, VDE, triggers a single double-strand break (DSB) at an inteinless allele, leading to VMA1 intein homing. Besides the accumulating information on the in vitro activity of VDE, very little has been known about the molecular mechanism of intein homing in yeast nucleus. RESULTS: We developed an assay to detect the product of VMA1 intein homing in yeast genome. We analysed mutant phenotypes of RecA homologs, Rad51p and Dmc1p, and their interacting proteins, Rad54p and Tid1p, and found that they all play critical roles in intein inheritance. The absence of DSB end processing proteins, Sae2p and those in the Mre11-Rad50-Xrs2 complex, also causes partial reduction in homing efficiency. As with meiotic recombination, crossover events are frequently observed during intein homing. We also observed that the absence of premeiotic DNA replication caused by hydroxyurea (HU) or clb5delta clb6delta mutation reduces VDE-mediated DSBs. CONCLUSION: The repairing system working in intein homing shares molecular machinery with meiotic recombination induced by Spo11p. Moreover, like Spo11p-induced DNA cleavage, premeiotic DNA replication is a prerequisite for a VDE-induced DSB. VMA1 intein thus utilizes several host factors involved in meiotic and recombinational processes to spread its genetic information and guarantee its progeny through establishment of a parasitic relationship with the organism.


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 Nucleic Acids ResHome page
T. Fukuda, K. Kugou, H. Sasanuma, T. Shibata, and K. Ohta
Targeted induction of meiotic double-strand breaks reveals chromosomal domain-dependent regulation of Spo11 and interactions among potential sites of meiotic recombination
Nucleic Acids Res., February 11, 2008; 36(3): 984 - 997.
[Abstract] [Full Text] [PDF]

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 Eukaryot CellHome page
T. Fukuda, K. Ohta, and Y. Ohya
Investigation of the Mechanism of Meiotic DNA Cleavage by VMA1-Derived Endonuclease Uncovers a Meiotic Alteration in Chromatin Structure around the Target Site
Eukaryot. Cell, June 1, 2006; 5(6): 981 - 990.
[Abstract] [Full Text] [PDF]

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 Nucleic Acids ResHome page
K. L. Posey, V. Koufopanou, A. Burt, and F. S. Gimble
Evolution of divergent DNA recognition specificities in VDE homing endonucleases from two yeast species
Nucleic Acids Res., July 27, 2004; 32(13): 3947 - 3956.
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