In sexually reproducing animals, a crucial step in zygote formation is the decondensation of the fertilizing sperm nucleus into a DNA replication-competent male pronucleus. Genome-wide nucleosome assembly on paternal DNA implies the replacement of sperm chromosomal proteins, such as protamines, by maternally provided histones1,2. This fundamental process is specifically impaired in sésame (ssm), a unique Drosophila maternal effect mutant that prevents male pronucleus formation3. Here we show that ssm is a point mutation in the Hira gene, thus demonstrating that the histone chaperone protein HIRA is required for nucleosome assembly during sperm nucleus decondensation. In vertebrates, HIRA has recently been shown to be critical for a nucleosome assembly pathway independent of DNA synthesis that specifically involves the H3.3 histone variant4,5. We also show that nucleosomes containing H3.3, and not H3, are specifically assembled in paternal Drosophila chromatin before the first round of DNA replication. The exclusive marking of paternal chromosomes with H3.3 represents a primary epigenetic distinction between parental genomes in the zygote, and underlines an important consequence of the critical and highly specialized function of HIRA at fertilization.
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We thank P. Fisher for anti-PCNA antibodies and the European Drosophila Genome Project for cosmid clones. We are grateful to B. Durand and B. Horard for helpful discussions. We also thank J. Schmitt and the CTµ microscopy center for technical assistance. This work was supported by the C.N.R.S. and the French Ministry of Research.
Reprints and permissions information is available at npg.nature.com/reprintsandpermissions. The authors declare no competing financial interests.
RT-PCR analysis of Hira expression (PDF 735 kb)
Western Blot analysis of HIRA-FLAG, H3.3-FLAG and H3-FLAG proteins. (PDF 283 kb)
Pronuclei replicate their DNA by the time they appose. (PDF 532 kb)
Complementation of the ssm185b phenotype with Hira transgenes. (PDF 783 kb)
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