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Molecular and Cellular Biology, February 2003, p. 1151-1162, Vol. 23, No. 4
0270-7306/03/$08.00+0     DOI: 10.1128/MCB.23.4.1151-1162.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Loss of HR6B Ubiquitin-Conjugating Activity Results in Damaged Synaptonemal Complex Structure and Increased Crossing-Over Frequency during the Male Meiotic Prophase

Willy M. Baarends,1* Evelyne Wassenaar,1 Jos W. Hoogerbrugge,1 Gert van Cappellen,1 Henk P. Roest,2 Jan Vreeburg,1 Marja Ooms,1 Jan H. J. Hoeijmakers,2 and J. Anton Grootegoed1

Department of Reproduction and Development,1 Department of Cell Biology and Genetics, Erasmus University Rotterdam, 3000 DR Rotterdam, The Netherlands2

Received 9 July 2002/ Returned for modification 9 August 2002/ Accepted 19 November 2002

The ubiquitin-conjugating enzymes HR6A and HR6B are the two mammalian homologs of Saccharomyces cerevisiae RAD6. In yeast, RAD6 plays an important role in postreplication DNA repair and in sporulation. HR6B knockout mice are viable, but spermatogenesis is markedly affected during postmeiotic steps, leading to male infertility. In the present study, increased apoptosis of HR6B knockout primary spermatocytes was detected during the first wave of spermatogenesis, indicating that HR6B performs a primary role during the meiotic prophase. Detailed analysis of HR6B knockout pachytene nuclei showed major changes in the synaptonemal complexes. These complexes were found to be longer. In addition, we often found depletion of synaptonemal complex proteins from near telomeric regions in the HR6B knockout pachytene nuclei. Finally, we detected an increased number of foci containing the mismatch DNA repair protein MLH1 in these nuclei, reflecting a remarkable and consistent increase (20 to 25%) in crossing-over frequency. The present findings reveal a specific requirement for the ubiquitin-conjugating activity of HR6B in relation to dynamic aspects of the synaptonemal complex and meiotic recombination in spermatocytes.

* Corresponding author. Mailing address: Department of Reproduction and Development, Erasmus University Rotterdam, P.O. Box 1738, 3000 DR Rotterdam, The Netherlands. Phone: 31-10-4087976. Fax: 31-10-4089461. E-mail: baarends{at}endov.fgg.eur.nl.

Molecular and Cellular Biology, February 2003, p. 1151-1162, Vol. 23, No. 4
0022-538X/03/$08.00+0     DOI: 10.1128/MCB.23.4.1151-1162.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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