Skip to main content
NCBI home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1990 Dec;10(12):6730–6741. doi: 10.1128/mcb.10.12.6730

Activation of a late H2B histone gene in blastula-stage sea urchin embryos by an unusual enhancer element located 3' of the gene.

A Z Zhao 1, A M Colin 1, J Bell 1, M Baker 1, B R Char 1, R Maxson 1
PMCID: PMC362951  PMID: 2247080

Abstract

In the sea urchin embryo, late histone genes are transcribed at low levels during cleavage and blastula formation and at substantially higher levels in later stages of embryogenesis. To investigate the molecular basis of the stage-specific expression of a late H2B histone gene, we injected mutant genes lacking portions of 5'- and 3'-flanking regions into Lytechinus pictus embryos and monitored their expression by RNase protection. A 200-bp region located 489 bp downstream of the mRNA 3' terminus was necessary for the increase in transcription of the late H2B gene at the mid-blastula stage of development. DNase I and methylation interference footprint analyses located only one factor-binding site in this region, and gel mobility shift experiments showed that the DNA-binding activity of this factor (designated H2B abp 1) paralleled the transcriptional activity of the L1 H2B gene. Additional mutagenesis and microinjection experiments located the activator element to a 32-bp DNA segment that includes the H2B abp 1-binding site. These experiments also showed that the 32-bp fragment functions independently of position and orientation and therefore has the hallmarks of an enhancer. That this fragment contains most or all of the L1 H2B gene transcription-stimulatory activity makes it unusual among enhancerlike elements, which generally consist of several clustered factor-binding sites that act additively or cooperatively to affect transcription. The nucleotide sequence of the L1 H2B enhancer element suggests that the trans-acting factor that interacts with it is a member of the antennapedia or engrailed class of homeodomain proteins.

Full text

PDF
6730

Images in this article

6732Image
on p.6732
6735Image
on p.6735
6736Image
on p.6736
6736Image
on p.6736
6737Image
on p.6737
6738Image
on p.6738

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Anderson K. V., Lengyel J. A. Changing rates of histone mRNA synthesis and turnover in Drosophila embryos. Cell. 1980 Oct;21(3):717–727. doi: 10.1016/0092-8674(80)90435-3. [DOI] [PubMed] [Google Scholar]
  2. Barberis A., Superti-Furga G., Busslinger M. Mutually exclusive interaction of the CCAAT-binding factor and of a displacement protein with overlapping sequences of a histone gene promoter. Cell. 1987 Jul 31;50(3):347–359. doi: 10.1016/0092-8674(87)90489-2. [DOI] [PubMed] [Google Scholar]
  3. Barberis A., Superti-Furga G., Vitelli L., Kemler I., Busslinger M. Developmental and tissue-specific regulation of a novel transcription factor of the sea urchin. Genes Dev. 1989 May;3(5):663–675. doi: 10.1101/gad.3.5.663. [DOI] [PubMed] [Google Scholar]
  4. Busslinger M., Barberis A. Synthesis of sperm and late histone cDNAs of the sea urchin with a primer complementary to the conserved 3' terminal palindrome: evidence for tissue-specific and more general histone gene variants. Proc Natl Acad Sci U S A. 1985 Sep;82(17):5676–5680. doi: 10.1073/pnas.82.17.5676. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Calzone F. J., Thézé N., Thiebaud P., Hill R. L., Britten R. J., Davidson E. H. Developmental appearance of factors that bind specifically to cis-regulatory sequences of a gene expressed in the sea urchin embryo. Genes Dev. 1988 Sep;2(9):1074–1088. doi: 10.1101/gad.2.9.1074. [DOI] [PubMed] [Google Scholar]
  6. Carroll S. B., Scott M. P. Localization of the fushi tarazu protein during Drosophila embryogenesis. Cell. 1985 Nov;43(1):47–57. doi: 10.1016/0092-8674(85)90011-x. [DOI] [PubMed] [Google Scholar]
  7. Childs G., Maxson R., Kedes L. H. Histone gene expression during sea urchin embryogenesis: isolation and characterization of early and late messenger RNAs of Strongylocentrotus purpuratus by gene-specific hybridization and template activity. Dev Biol. 1979 Nov;73(1):153–173. doi: 10.1016/0012-1606(79)90144-1. [DOI] [PubMed] [Google Scholar]
  8. Childs G., Nocente-McGrath C., Lieber T., Holt C., Knowles J. A. Sea urchin (lytechinus pictus) late-stage histone H3 and H4 genes: characterization and mapping of a clustered but nontandemly linked multigene family. Cell. 1982 Dec;31(2 Pt 1):383–393. doi: 10.1016/0092-8674(82)90132-5. [DOI] [PubMed] [Google Scholar]
  9. Chodosh L. A., Baldwin A. S., Carthew R. W., Sharp P. A. Human CCAAT-binding proteins have heterologous subunits. Cell. 1988 Apr 8;53(1):11–24. doi: 10.1016/0092-8674(88)90483-7. [DOI] [PubMed] [Google Scholar]
  10. Colin A. M., Catlin T. L., Kidson S. H., Maxson R. Closely linked early and late histone H2B genes are differentially expressed after microinjection into sea urchin zygotes. Proc Natl Acad Sci U S A. 1988 Jan;85(2):507–510. doi: 10.1073/pnas.85.2.507. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Colin A. M. Rapid repetitive microinjection. Methods Cell Biol. 1986;27:395–406. [PubMed] [Google Scholar]
  12. Cox K. H., Angerer L. M., Lee J. J., Davidson E. H., Angerer R. C. Cell lineage-specific programs of expression of multiple actin genes during sea urchin embryogenesis. J Mol Biol. 1986 Mar 20;188(2):159–172. doi: 10.1016/0022-2836(86)90301-3. [DOI] [PubMed] [Google Scholar]
  13. Desplan C., Theis J., O'Farrell P. H. The sequence specificity of homeodomain-DNA interaction. Cell. 1988 Sep 23;54(7):1081–1090. doi: 10.1016/0092-8674(88)90123-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. DiLiberto M., Lai Z. C., Fei H., Childs G. Developmental control of promoter-specific factors responsible for the embryonic activation and inactivation of the sea urchin early histone H3 gene. Genes Dev. 1989 Jul;3(7):973–985. doi: 10.1101/gad.3.7.973. [DOI] [PubMed] [Google Scholar]
  15. Dolecki G. J., Wang G., Humphreys T. Stage- and tissue-specific expression of two homeo box genes in sea urchin embryos and adults. Nucleic Acids Res. 1988 Dec 23;16(24):11543–11558. doi: 10.1093/nar/16.24.11543. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Fjose A., McGinnis W. J., Gehring W. J. Isolation of a homoeo box-containing gene from the engrailed region of Drosophila and the spatial distribution of its transcripts. Nature. 1985 Jan 24;313(6000):284–289. doi: 10.1038/313284a0. [DOI] [PubMed] [Google Scholar]
  17. Flytzanis C. N., Britten R. J., Davidson E. H. Ontogenic activation of a fusion gene introduced into sea urchin eggs. Proc Natl Acad Sci U S A. 1987 Jan;84(1):151–155. doi: 10.1073/pnas.84.1.151. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Fried M., Crothers D. M. Equilibria and kinetics of lac repressor-operator interactions by polyacrylamide gel electrophoresis. Nucleic Acids Res. 1981 Dec 11;9(23):6505–6525. doi: 10.1093/nar/9.23.6505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Galas D. J., Schmitz A. DNAse footprinting: a simple method for the detection of protein-DNA binding specificity. Nucleic Acids Res. 1978 Sep;5(9):3157–3170. doi: 10.1093/nar/5.9.3157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Gray H. B., Jr, Ostrander D. A., Hodnett J. L., Legerski R. J., Robberson D. L. Extracellular nucleases of Pseudomonas BAL 31. I. Characterization of single strand-specific deoxyriboendonuclease and double-strand deoxyriboexonuclease activities. Nucleic Acids Res. 1975 Sep;2(9):1459–1492. doi: 10.1093/nar/2.9.1459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hafen E., Kuroiwa A., Gehring W. J. Spatial distribution of transcripts from the segmentation gene fushi tarazu during Drosophila embryonic development. Cell. 1984 Jul;37(3):833–841. doi: 10.1016/0092-8674(84)90418-5. [DOI] [PubMed] [Google Scholar]
  22. Halsell S. R., Ito M., Maxson R. Differential expression of early and late embryonic histone genes in adult tissues of the sea urchin Strongylocentrotus purpuratus. Dev Biol. 1987 Jan;119(1):268–274. doi: 10.1016/0012-1606(87)90228-4. [DOI] [PubMed] [Google Scholar]
  23. Heberlein U., England B., Tjian R. Characterization of Drosophila transcription factors that activate the tandem promoters of the alcohol dehydrogenase gene. Cell. 1985 Jul;41(3):965–977. doi: 10.1016/s0092-8674(85)80077-5. [DOI] [PubMed] [Google Scholar]
  24. Heberlein U., Tjian R. Temporal pattern of alcohol dehydrogenase gene transcription reproduced by Drosophila stage-specific embryonic extracts. Nature. 1988 Feb 4;331(6155):410–415. doi: 10.1038/331410a0. [DOI] [PubMed] [Google Scholar]
  25. Hendrickson W., Schleif R. A dimer of AraC protein contacts three adjacent major groove regions of the araI DNA site. Proc Natl Acad Sci U S A. 1985 May;82(10):3129–3133. doi: 10.1073/pnas.82.10.3129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Hough-Evans B. R., Britten R. J., Davidson E. H. Mosaic incorporation and regulated expression of an exogenous gene in the sea urchin embryo. Dev Biol. 1988 Sep;129(1):198–208. doi: 10.1016/0012-1606(88)90174-1. [DOI] [PubMed] [Google Scholar]
  27. Hough-Evans B. R., Franks R. R., Cameron R. A., Britten R. J., Davidson E. H. Correct cell-type-specific expression of a fusion gene injected into sea urchin eggs. Dev Biol. 1987 Jun;121(2):576–579. doi: 10.1016/0012-1606(87)90193-x. [DOI] [PubMed] [Google Scholar]
  28. Ito M., Bell J., Lyons G., Maxson R. Synthesis and turnover of late H2B histone mRNA in developing embryos of the sea urchin, Strongylocentrotus purpuratus. Dev Biol. 1988 Sep;129(1):147–158. doi: 10.1016/0012-1606(88)90169-8. [DOI] [PubMed] [Google Scholar]
  29. Jaynes J. B., O'Farrell P. H. Activation and repression of transcription by homoeodomain-containing proteins that bind a common site. Nature. 1988 Dec 22;336(6201):744–749. doi: 10.1038/336744a0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Kaumeyer J. F., Weinberg E. S. Sequence, organization and expression of late embryonic H3 and H4 histone genes from the sea urchin, Strongylocentrotus purpuratus. Nucleic Acids Res. 1986 Jun 11;14(11):4557–4576. doi: 10.1093/nar/14.11.4557. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Kedes L. H. Histone genes and histone messengers. Annu Rev Biochem. 1979;48:837–870. doi: 10.1146/annurev.bi.48.070179.004201. [DOI] [PubMed] [Google Scholar]
  32. Kemler I., Busslinger M. Characterization of two nonallelic pairs of late histone H2A and H2B genes of the sea urchin: differential regulation in the embryo and tissue-specific expression in the adult. Mol Cell Biol. 1986 Nov;6(11):3746–3754. doi: 10.1128/mcb.6.11.3746. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Knowles J. A., Childs G. J. Temporal expression of late histone messenger RNA in the sea urchin Lytechinus pictus. Proc Natl Acad Sci U S A. 1984 Apr;81(8):2411–2415. doi: 10.1073/pnas.81.8.2411. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Ko H. S., Fast P., McBride W., Staudt L. M. A human protein specific for the immunoglobulin octamer DNA motif contains a functional homeobox domain. Cell. 1988 Oct 7;55(1):135–144. doi: 10.1016/0092-8674(88)90016-5. [DOI] [PubMed] [Google Scholar]
  35. Kornberg T., Sidén I., O'Farrell P., Simon M. The engrailed locus of Drosophila: in situ localization of transcripts reveals compartment-specific expression. Cell. 1985 Jan;40(1):45–53. doi: 10.1016/0092-8674(85)90307-1. [DOI] [PubMed] [Google Scholar]
  36. Lai Z. C., DeAngelo D. J., DiLiberto M., Childs G. An embryonic enhancer determines the temporal activation of a sea urchin late H1 gene. Mol Cell Biol. 1989 Jun;9(6):2315–2321. doi: 10.1128/mcb.9.6.2315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Lai Z. C., Maxson R., Childs G. Both basal and ontogenic promoter elements affect the timing and level of expression of a sea urchin H1 gene during early embryogenesis. Genes Dev. 1988 Feb;2(2):173–183. doi: 10.1101/gad.2.2.173. [DOI] [PubMed] [Google Scholar]
  38. Lieber T., Weisser K., Childs G. Analysis of histone gene expression in adult tissues of the sea urchins Strongylocentrotus purpuratus and Lytechinus pictus: tissue-specific expression of sperm histone genes. Mol Cell Biol. 1986 Jul;6(7):2602–2612. doi: 10.1128/mcb.6.7.2602. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Livant D. L., Cutting A. E., Britten R. J., Davidson E. H. An in vivo titration of regulatory factors required for expression of a fusion gene in transgenic sea urchin embryos. Proc Natl Acad Sci U S A. 1988 Oct;85(20):7607–7611. doi: 10.1073/pnas.85.20.7607. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  41. Maxson R. E., Jr, Wilt F. H. Accumulation of the early histone messenger RNAs during the development of Strongylocentrotus purpuratus. Dev Biol. 1982 Dec;94(2):435–440. doi: 10.1016/0012-1606(82)90360-8. [DOI] [PubMed] [Google Scholar]
  42. Maxson R. E., Jr, Wilt F. H. The rate of synthesis of histone mRNA during the development of sea urchin embryos (Strongylocentrotus purpuratus). Dev Biol. 1981 Apr 30;83(2):380–386. doi: 10.1016/0012-1606(81)90485-1. [DOI] [PubMed] [Google Scholar]
  43. Maxson R., Cohn R., Kedes L., Mohun T. Expression and organization of histone genes. Annu Rev Genet. 1983;17:239–277. doi: 10.1146/annurev.ge.17.120183.001323. [DOI] [PubMed] [Google Scholar]
  44. Maxson R., Ito M., Balcells S., Thayer M., French M., Lee F., Etkin L. Differential stimulation of sea urchin early and late H2B histone gene expression by a gastrula nuclear extract after injection into Xenopus laevis oocytes. Mol Cell Biol. 1988 Mar;8(3):1236–1246. doi: 10.1128/mcb.8.3.1236. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Maxson R., Mohun T., Gormezano G., Childs G., Kedes L. Distinct organizations and patterns of expression of early and late histone gene sets in the sea urchin. Nature. 1983 Jan 13;301(5896):120–125. doi: 10.1038/301120a0. [DOI] [PubMed] [Google Scholar]
  46. Maxson R., Mohun T., Gormezano G., Kedes L. Evolution of late H2A, H2B, and H4 histone genes of the sea urchin, Strongylocentrotus purpuratus. Nucleic Acids Res. 1987 Dec 23;15(24):10569–10582. doi: 10.1093/nar/15.24.10569. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. McMahon A. P., Flytzanis C. N., Hough-Evans B. R., Katula K. S., Britten R. J., Davidson E. H. Introduction of cloned DNA into sea urchin egg cytoplasm: replication and persistence during embryogenesis. Dev Biol. 1985 Apr;108(2):420–430. doi: 10.1016/0012-1606(85)90045-4. [DOI] [PubMed] [Google Scholar]
  48. Melton D. A., Krieg P. A., Rebagliati M. R., Maniatis T., Zinn K., Green M. R. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 1984 Sep 25;12(18):7035–7056. doi: 10.1093/nar/12.18.7035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Müeller-Storm H. P., Sogo J. M., Schaffner W. An enhancer stimulates transcription in trans when attached to the promoter via a protein bridge. Cell. 1989 Aug 25;58(4):767–777. doi: 10.1016/0092-8674(89)90110-4. [DOI] [PubMed] [Google Scholar]
  50. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Schirm S., Jiricny J., Schaffner W. The SV40 enhancer can be dissected into multiple segments, each with a different cell type specificity. Genes Dev. 1987 Mar;1(1):65–74. doi: 10.1101/gad.1.1.65. [DOI] [PubMed] [Google Scholar]
  52. Siebenlist U., Gilbert W. Contacts between Escherichia coli RNA polymerase and an early promoter of phage T7. Proc Natl Acad Sci U S A. 1980 Jan;77(1):122–126. doi: 10.1073/pnas.77.1.122. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Sucov H. M., Hough-Evans B. R., Franks R. R., Britten R. J., Davidson E. H. A regulatory domain that directs lineage-specific expression of a skeletal matrix protein gene in the sea urchin embryo. Genes Dev. 1988 Oct;2(10):1238–1250. doi: 10.1101/gad.2.10.1238. [DOI] [PubMed] [Google Scholar]
  54. Sures I., Lowry J., Kedes L. H. The DNA sequence of sea urchin (S. purpuratus) H2A, H2B and H3 histone coding and spacer regions. Cell. 1978 Nov;15(3):1033–1044. doi: 10.1016/0092-8674(78)90287-8. [DOI] [PubMed] [Google Scholar]
  55. Vitelli L., Kemler I., Lauber B., Birnstiel M. L., Busslinger M. Developmental regulation of micro-injected histone genes in sea urchin embryos. Dev Biol. 1988 May;127(1):54–63. doi: 10.1016/0012-1606(88)90188-1. [DOI] [PubMed] [Google Scholar]
  56. Wang J. C., Giaever G. N. Action at a distance along a DNA. Science. 1988 Apr 15;240(4850):300–304. doi: 10.1126/science.3281259. [DOI] [PubMed] [Google Scholar]
  57. Weinberg E. S., Hendricks M. B., Hemminki K., Kuwabara P. E., Farrelly L. A. Timing and rates of synthesis of early histone mRNA in the embryo of Strongylocentrotus purpuratus. Dev Biol. 1983 Jul;98(1):117–129. doi: 10.1016/0012-1606(83)90340-8. [DOI] [PubMed] [Google Scholar]
  58. Weir M. P., Kornberg T. Patterns of engrailed and fushi tarazu transcripts reveal novel intermediate stages in Drosophila segmentation. Nature. 1985 Dec 5;318(6045):433–439. doi: 10.1038/318433a0. [DOI] [PubMed] [Google Scholar]
  59. Winslow G. M., Hayashi S., Krasnow M., Hogness D. S., Scott M. P. Transcriptional activation by the Antennapedia and fushi tarazu proteins in cultured Drosophila cells. Cell. 1989 Jun 16;57(6):1017–1030. doi: 10.1016/0092-8674(89)90340-1. [DOI] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES