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Mol Cell Biol, July 1998, p. 4337-4346, Vol. 18, No. 7
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Extensive, Nonrandom Diversity of Excision Footprints Generated by Ds-Like Transposon Ascot-1 Suggests New Parallels with V(D)J Recombination

Vincent Colot,* Vicki Haedens, and Jean-Luc Rossignoldagger

Institut de Génétique et Microbiologie, Centre National de la Recherche Scientifique-Unité de Recherche Associée 2225, Université Paris-Sud, F-91405 Orsay cedex, France

Received 29 January 1998/Returned for modification 13 March 1998/Accepted 2 April 1998

Upon insertion, transposable elements can disrupt or alter gene function in various ways. Transposons moving through a cut-and-paste mechanism are in addition often mutagenic when excising because repair of the empty site seldom restores the original sequence. The characterization of numerous excision events in many eukaryotes indicates that transposon excision from a given site can generate a high degree of DNA sequence and phenotypic variation. Whether such variation is generated randomly remains largely to be determined. To this end, we have exploited a well-characterized system of genetic instability in the fungus Ascobolus immersus to perform an extensive study of excision events. We show that this system, which produces many phenotypically and genetically distinct derivatives, results from the excision of a novel Ds-like transposon, Ascot-1, from the spore color gene b2. A unique set of 48 molecularly distinct excision products were readily identified from a representative sample of excision derivatives. Products varied in their frequency of occurrence over 4 orders of magnitude, yet most showed small palindromic nucleotide additions. Based on these and other observations, compelling evidence was obtained for intermediate hairpin formation during the excision reaction and for strong biases in the subsequent processing steps at the empty site. Factors likely to be involved in these biases suggest new parallels between the excision reaction performed by transposons of the hAT family and V(D)J recombination. An evaluation of the contribution of small palindromic nucleotide additions produced by transposon excision to the spectrum of spontaneous mutations is also presented.

* Corresponding author. Present address: Institut Jacques Monod, Centre National de la Recherche Scientifique-Unité Mixte de Recherche 7592, Département de Microbiologie, Universités Paris 6 et 7, 2 Place Jussieu, 75251 Paris cedex 05, France. Phone: (33) 1 44 27 40 95. Fax: (33) 1 44 27 82 10. E-mail: colot{at}ijm.jussieu.fr.

dagger Present address: Institut Jacques Monod, CNRS-UMR 7592, Département de Microbiologie, Universités Paris 6 et 7, 75251 Paris cedex 05, France.

Mol Cell Biol, July 1998, p. 4337-4346, Vol. 18, No. 7
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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