Conservative segregation of maternally inherited CS histone variants in larval stages of sea urchin development (original) (raw)
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Nucleic Acids Research, 1979
Histone mRNAs at different stages of development were purified by hybridization with the cloned homologous histone genes. The electrophoretic patterns of oocytes, 2-4 blastomeres, 64 cells and morula histone mRNAs was found to be identical, whereas the electrophoretic pattern of mesenchyme blastula histone mRNA was aseddebr different. The cloned histone DNA of P.lividus was hybridized with the RNA of each stage. The Tm was 74°C in all cases except for the mesenchyme histone mRNAs whose Tm was 59°C, thus suggesting that at least two different clusters of histone genes are active in the course of the sea urchin development.
Microheterogeneity of late histones in larval stages of sea urchin development
Comparative Biochemistry and Physiology Part B: Comparative Biochemistry, 1990
The total histone complement of early plutei were compared with that of intermediate and late larvae of the sea urchin Tefrapygus niger. 2. Electrophoretic comparison indicates that there are quantitative and qualitative shifts of the five classes throughout late larval development. 3. The strong similarity in the amino acid composition of total histones isolated from early, intermediate and late plutei indicates that the observed electrophoretic heterogeneity is due to post-translational modifications.
Molecular and cellular biology, 1981
We have examined histone gene expression during the early stages of sea urchin embryogenesis. The five histone genes expressed at that time are contained in tandem repetitive segments. It has been suggested that adjacent coding regions and their intervening spacer sequences are transcribed into large polycistronic messenger ribonucleic acid (RNA) precursors. We have subcloned into pBR322 deoxyribonucleic acid (DNA) sequences mapping either in the coding region, the 5' spacer, or the 3' spacer of the H2B histone gene. These clones were used to produce radioiodinated hybridization probes. We measured the steady-state quantity of H2B messenger RNA as well as spacer-specific RNA in the total RNA from embryos taken at various stages of development from fertilization to hatching of blastulae (0 to 22 h post-fertilization). Small amounts of RNA hybridizing to both spacer probes could be found. However, we show that these RNAs form mismatched hybrids with the spacer DNA and therefor...
Chromatin structure of histone genes in sea urchin sperms and embryos
Nucleic Acids Research, 1982
The nucleosomal organization of active and repressed a subtype histone genes has been investigated by micrococcal nuclease digestion of P.lividus sperm, 32-64 S11 embryo and mesenchyme blastula nuclei, followed by hybridization with P-labeled specific DNA probes. In sperms, fully repressed histone genes are regularly folded in nucleosomes, and exhibit a greater resistance to micrococcal nuclease cleavage than bulk chromatin. In contrast, both coding and spacer a subtype histone DNA sequences acquire an altered conformation in nuclei from early cleavage stage embryos, i.e., when these genes are maximally expressed. Switching off of the a subtype histone genes, in mesenchyme blastulae, restores the typical nucleosomal organization on this chromatin region. As probed by hybridization to D.melanogaster actin cDNA, actin genes retain a regular nucleosomal structure in all the investigated stages.
Developmental Biology, 1987
The sea urchin synthesizes distinct classes of histone mRNAs at different stages of development. "Early" embryonic histone mRNAs are synthesized in large amount in cleavage and blastula stage embryos. "Late" embryonic histone mRNAs are the predominant forms in postblastula embryos. To learn more about how early and late histone genes are regulated during the life cycle of the sea urchin and to search for additional classes of developmentally regulated histone mRNAs, we examined histone mRNAs in sea urchin adult tissues. Using methods of primer extension and Sl nuclease protection, we found that tube foot, intestine, testis, and ovary contain a subset of the several H2b mRNA species synthesized by the embryo. We detected early H2b mRNA in ovary, but not. in other tissues. Three late H2b mRNA species were present in all tissues tested, while a fourth late H2b was not detected. Using a probe that hybridized specifically with transcripts of a single-copy late H2b gene, we found that this gene was transcribed in both embryos and adults. Interestingly, its level of expression relative to other late H2b genes varied among tissues. Finally, we identified two H2b mRNA species that were distinct from early and late embryonic forms and were synthesized only in adult tissues. 0 1987 Academic PWSS. I~C.
Histone phosphorylation during sea urchin development
Seminars in Cell Biology, 1995
Studies on histone phosphorylation during transitions in chromatin structure occurring in vivo during spermatogenesis and early embryogenesis in sea urchins are reviewed and evaluated in the light of recent studies on histone phosphorylation occurring during chromatin synthesis in frog egg extracts in vitro and evidence that protein kinases and phosphatases play direct roles in the regulation of cellular structure. Sperm-specific histone variants Sp H1 and Sp H2B are maintained as phosphorylated derivatives N and O/P throughout spermatogenesis and early embryogenesis and egg specific histone variants CS H1 and CS H2A are phosphorylated during early embryogenesis. These developmental correlations provide clues about the roles of histone phosphorylation in control of chromatin structure in vivo and provide a basis for the interpretation of data obtained from in-vitro sperm chromatin remodeling in egg extracts and from biochemical studies on the effects of histone phosphorylation on DNA binding. The potential consequences for chromatin structure of the various histone phosphorylation events observed in sea urchins and frog egg extracts are discussed.
European Journal of Biochemistry, 1997
The cleavage stage (CS) H1, H2A, and H2B histones of the sea urchin, which have previously been identified by their distinct electrophoretic mobility on Tritonlacidurea gels, are known to be maternally expressed during oogenesis and have been implicated in chromatin remodeling of the male pronucleus following fertilization. Here, we describe the isolation of these three CS histones by reverse-phase HPLC chromatography. Moreover, a novel CS H3 protein was identified by the same purification procedure. A low incorporation of radioactive amino acids into the CS hiqtones during early development revealed that the bulk of these proteins in the blastula embryo are derived from the maternal pool of the egg. Amino acid analysis, together with the previously described electrophoretic mobilities, unequivocally identified the purified proteins as CS histones. Peptide sequence analysis confirmed the novel nature of the CS variants as they are distantly related to the early, late, and sperm histone subtypes of the sea urchin. The CS H1 protein displays highest sequence similarity with the H1M (B4) histone of Xenopus laevis, indicating that the frog H1M protein may be a vertebrate homologue of the CS HI histone. These data suggest an ancient evolutionary origin and wide distribution of the CS histone variants.
Embryonic regulation of histone ubiquitination the sea urchin
Developmental Genetics, 1995
We have used quantitative 2-D protein electrophoresis and immunoprecipitation to study the patterns of histone ubiquitination at 10 h and 36 h of embryonic development in Strongylocentrotus purpuratus. Variants csH2A, aH2A, PH2A, yH2A, SHA, H2AF./Z, aHZB, pH26, and yH2B showed up to sevenfold differences in level of monoubiquitination between variants, and individual variants showed up to sixfold changes during development. At 36 h of embryogenesis, the late variants were less ubiquitinated than the early variants, a I thoug h the overa II level of u big u itina tion was appreciably greater than at 10 h. Antiubiquitin antibodies were used to precipitate formaldehydefixed chromatin fragments in order to estimate the degree of ubiquitination of the early histone genes.