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First published online 8 May 2007
doi: 10.1242/jcs.03451

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Increased phosphorylation and dimethylation of XY body histones in the Hr6b-knockout mouse is associated with derepression of the X chromosome

¹ Department of Reproduction and Development, Erasmus MC–University Medical Center, Rotterdam, The Netherlands
² Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, TN, USA

^* Author for correspondence (e-mail: w.baarends{at}erasmusmc.nl)

Accepted 4 April 2007

Mono-ubiquitylated H2A marks the transcriptionally silenced XY body during male meiotic prophase. Concomitant with H2A_K119ub1, the ubiquitin-conjugating enzyme HR6B is also enriched on the XY body. We analyzed H2A and H2B ubiquitylation in Hr6b-knockout mouse spermatocytes, but no global changes were detected. Next, we analyzed phosphorylation of the threonine residues T120 and T119 that are adjacent to the K119 and K120 target sites for ubiquitylation in H2A and H2B, respectively. In wild-type cells, H2A_T120ph and H2B_T119ph mark meiotically unpaired and silenced chromatin, including the XY body. In Hr6b-knockout spermatocytes, the H2B_T119ph signal was unchanged, but H2A_T120ph was enhanced from late pachytene until metaphase I. Furthermore, we found increased H3_K4 dimethylation on the X and Y chromosomes of diplotene Hr6b-knockout spermatocytes, persisting into postmeiotic round spermatids. In these cells, the X and Y chromosomes maintained an unchanged H3_K9m2 level, even when this modification was lost from centromeric heterochromatin. Analysis of gene expression showed derepression of X chromosome genes in postmeiotic Hr6b-knockout spermatids. We conclude that HR6B exerts control over different histone modifications in spermatocytes and spermatids, and that this function contributes to the postmeiotic maintenance of X chromosome silencing.

Key words: RAD6, HR6A, HR6B, Meiosis, XY body, Histone modifications

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