Mouse sperm chromatin proteins: quantitative isolation and partial characterization

Balhorn, Rod; Gledhill, Barton L.; Wyrobek, Andrew J.

doi:10.1021/bi00637a021

Cited by 225 publications

(140 citation statements)

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“…In many metazoans, male germ cells undergo during their final differentiation into sperm an extensive chromatin remodeling process during which genomic DNA becomes newly packaged into a highly condensed configuration by sperm specific proteins. In mammals, removal of histones is generally not complete 10,11,[19][20][21][22][23][24] . Furthermore, remaining histones have been reported to stay associated with the paternal genome during de novo chromatin formation in the zygote following fertilization 9 .…”

mentioning

confidence: 99%

Molecular determinants of nucleosome retention at CpG-rich sequences in mouse spermatozoa

Erkek

Hisano

Liang

et al. 2013

Nat Struct Mol Biol

302

379

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In mammals, fusion of two morphologically distinct gametes, oocytes and spermatozoa, leads to the formation of totipotent embryos. Acquisition of totipotency is thought to be mediated by extensive epigenetic reprogramming of parental genomes, affecting DNA methylation and histone modifications, and possibly replication timing and transcriptional activity in parental specific manners [1][2][3][4] . It is currently unclear to what extent differential reprogramming of maternal and paternal genomes is due to differences in chromatin states inherited from the oocyte and spermatozoon [4][5][6][7][8][9][10][11] . Beyond DNA methylation 1,2,6,12 , it is unknown which types of parental chromatin states are maintained or reprogrammed in early embryos. If certain parental chromatin states did escape reprogramming in the early embryo, such information could constitute an "intrinsic intergenerational epigenetic program directing gene expression in the next generation 13 . If these chromatin states also escaped reprogramming during gametogenesis, the inheritance program would function transgenerationally 13 . An increasing number of studies point to inter-or transgenerational transmission of acquired phenotypic traits that are related to temporal exposure of (grand-)parents to alternative instructive environmental cues [14][15][16][17][18] . Mechanistically, such phenotypic changes may be related to (transient) alterations of an intrinsic inheritance program.A role of histones and associated posttranslational modifications in maternal and paternal transmission of intrinsic or acquired epigenetic information is largely unknown 13 . In many metazoans, male germ cells undergo during their final differentiation into sperm an extensive chromatin remodeling process during which 3 genomic DNA becomes newly packaged into a highly condensed configuration by sperm specific proteins. In mammals, removal of histones is generally not complete 10,11,[19][20][21][22][23][24] . Furthermore, remaining histones have been reported to stay associated with the paternal genome during de novo chromatin formation in the zygote following fertilization 9 .We and others recently showed that histones lasting in human sperm are to some extent enriched at regulatory sequences of genes 10,11 . We also demonstrated that H3K4me3-and H3K27me3-marked histones are retained at promoters of specific sets of genes in mouse spermatozoa 11. The extent of evolutionary conservation of nucleosome retention at gene regulatory sequences in spermatozoa and the mechanistic principles of such retention are, however, unknown.To address conservation and to dissect the molecular logic underlying nucleosome retention, we determined the genome-wide nucleosome occupancy in mouse spermatozoa that only contain 1% residual histones. We show here that combinatorial effects of sequence composition, histone variants and histone modifications uniquely determine the packaging of sperm DNA. Nucleosomes in sperm mainly localize to unmethylated CpG-rich sequences in a histone variant specif...

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mentioning

confidence: 99%

Molecular determinants of nucleosome retention at CpG-rich sequences in mouse spermatozoa

Erkek

Hisano

Liang

et al. 2013

Nat Struct Mol Biol

302

379

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“…The sperm were subsequently washed two times in Tris-saline with brief sonication (Sonifier; Branson Sonic Power Co., Danbury, Conn.; setting 1 for 5 to 10 s) and pelleted by centrifugation at 3,000 x g for 3 min. The spermatids and sperm were reduced with 10 mM dithiothreitol-0.01 M Tris (pH 8.0), and the tails, acrosomes, and membranes were removed by treatment with 1% mixed alkyltrimethylammonium bromide (5). The resulting amembranous nuclei were washed in Tris-saline and dissolved in 5 M guanidine hydrochloride, and the basic proteins were extracted as described previously (5).…”

Section: Methodsmentioning

confidence: 99%

Mouse protamine 2 is synthesized as a precursor whereas mouse protamine 1 is not.

Yelick¹,

Balhorn²,

Johnson³

et al. 1987

Mol. Cell. Biol.

147

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The nuclei of mouse spermatozoa contain two protamine variants, mouse protamine 1 (mPl) and mouse protamine 2 (mP2). The amino acid sequence predicted from mPl cDNAs demonstrates that mPl is a 50-amino-acid protein with strong homology to other mammalian P1 protamines. Nucleotide sequence analysis of independently isolated, overlapping cDNA clones indicated that mP2 is initially synthesized as a precursor protein which is subsequently processed into the spermatozoan form of mP2. The existence of the mP2 precursor was confirmed by amino acid composition and sequence analysis of the largest of a set of four basic proteins isolated from late-step spermatids whose synthesis is coincident with that of mPl. The sequence of the first 10 amino acids of this protein, mP2 precursor 1, exactly matches that predicted from the nucleotide sequence of cDNA and genomic mP2 clones. The amino acid composition of isolated mP2 precursor 1 very closely matches that predicted from the mP2 cDNA nucleotide sequence. Sequence analysis of the amino terminus of isolated mature mP2 identified the final processing point within the mP2 precursor. These studies demonstrated that mP2 is synthesized as a precursor containing 106 amino acids which is processed into the mature, 63-amino-acid form found in spermatozoa.Protamines are small, basic, arginine-rich proteins that replace histones in the later stages of spermatogenesis in many vertebrates (N. B. Hecht, in G. Stein and J. Stein, ed., Basic Chromosomal Proteins: Structure, Organization and Regulation ofthe Gene, in press). During spermatogenesis in mammals, the histones are not directly replaced by protamines but by a population of basic proteins that are transiently associated with the spermatid nucleus (14,35; Hecht, in press). These testis-specific proteins are removed from the condensing chromatin at later stages of spermiogenesis and are replaced by protamines, the predominant class of proteins found complexed with DNA in the mature spermatozoon. Mammalian protamines contain the amino acid cysteine, which is believed to function in the stabilization and compaction of the sperm nucleus through disulfide bond cross-linking (7).Protamines have been isolated from a large number of vertebrates including fish (3, 28), domestic fowl (38), mice (6, 27, 40; A. R.

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“…Following meiosis, haploid round spermatids undergo dramatic and extensive chromatin remodeling. This process results in the genome-wide exchange of histones by spermatogenesis-specific basic DNA packaging proteins (initially transition proteins and ultimately protamines) [64], [65].…”

Section: Global Chromatin Remodeling During Spermiogenesismentioning

confidence: 99%

Parental epigenetic control of embryogenesis: a balance between inheritance and reprogramming?

Gill

Erkek

Peters

2012

Current Opinion in Cell Biology

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At fertilization, fusion of two differentiated gametes forms the zygote that is capable of forming all of the varied cell lineages of an organism. It is widely thought that the acquisition of totipotency involves extensive epigenetic reprogramming of the germline state into an embryonic state. However, recent data argue that this reprogramming is incomplete and that substantial epigenetic information passes from one generation to the next. In this review we summarize the changes in chromatin states that take place during mammalian gametogenesis and examine the evidence that early mammalian embryogenesis may be affected by inheritance of epigenetic information from the parental generation.

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Mouse sperm chromatin proteins: quantitative isolation and partial characterization

Cited by 225 publications

References 22 publications

Molecular determinants of nucleosome retention at CpG-rich sequences in mouse spermatozoa

Molecular determinants of nucleosome retention at CpG-rich sequences in mouse spermatozoa

Mouse protamine 2 is synthesized as a precursor whereas mouse protamine 1 is not.

Parental epigenetic control of embryogenesis: a balance between inheritance and reprogramming?

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