Skip to main content
PMC home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1989 May 11;17(9):3563–3582. doi: 10.1093/nar/17.9.3563

Multiple regulatory elements of the murine gamma 2-crystallin promoter.

S Lok 1, W Stevens 1, M L Breitman 1, L C Tsui 1
PMCID: PMC317796  PMID: 2726487

Abstract

Crystallins are the major water-soluble proteins of the vertebrate eye lens. These lens-specific proteins are encoded by several multi-gene families whose expression is differentially regulated during development. Our previous studies showed that the mouse gamma 2-crystallin promoter is active on transfection into lens-explant cultures derived from 14-day-old chick embryos but not on transfection into a variety of non-lens cells. In this study, transient expression data show that a sequence of 226 nucleotides upstream from the transcription start site is sufficient for activity of this promoter in the chicken lens cells. This sequence can be further divided into two domains, A and B, both of which are required for promoter function. Domain A (nucleotide -68 to -18) contains the TATA box and sequence motifs that are conserved in all gamma-crystallin promoters. Domain B (-226 to -120) consists of three regions. One of these regions contains an element with dyad symmetry and a sequence similar to the octamer motif. The second region contains an enhancer core consensus sequence. Two "enhancer-like" activities have been detected, one in Domain B and a second in a more distal region (-392 to -278) that does not appear to be required for promoter activity in transfection assays.

Full text

PDF
3566

Images in this article

3569Image
on p.3569
3576Image
on p.3576

Selected References

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

  1. Baumruker T., Sturm R., Herr W. OBP100 binds remarkably degenerate octamer motifs through specific interactions with flanking sequences. Genes Dev. 1988 Nov;2(11):1400–1413. doi: 10.1101/gad.2.11.1400. [DOI] [PubMed] [Google Scholar]
  2. Beato M. Gene regulation by steroid hormones. Cell. 1989 Feb 10;56(3):335–344. doi: 10.1016/0092-8674(89)90237-7. [DOI] [PubMed] [Google Scholar]
  3. Bloemendal H. The vertebrate eye lens. Science. 1977 Jul 8;197(4299):127–138. doi: 10.1126/science.877544. [DOI] [PubMed] [Google Scholar]
  4. Borrás T., Nickerson J. M., Chepelinsky A. B., Piatigorsky J. Structural and functional evidence for differential promoter activity of the two linked delta-crystallin genes in the chicken. EMBO J. 1985 Feb;4(2):445–452. doi: 10.1002/j.1460-2075.1985.tb03649.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Breitman M. L., Lok S., Wistow G., Piatigorsky J., Tréton J. A., Gold R. J., Tsui L. C. Gamma-crystallin family of the mouse lens: structural and evolutionary relationships. Proc Natl Acad Sci U S A. 1984 Dec;81(24):7762–7766. doi: 10.1073/pnas.81.24.7762. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Briggs M. R., Kadonaga J. T., Bell S. P., Tjian R. Purification and biochemical characterization of the promoter-specific transcription factor, Sp1. Science. 1986 Oct 3;234(4772):47–52. doi: 10.1126/science.3529394. [DOI] [PubMed] [Google Scholar]
  7. Chepelinsky A. B., King C. R., Zelenka P. S., Piatigorsky J. Lens-specific expression of the chloramphenicol acetyltransferase gene promoted by 5' flanking sequences of the murine alpha A-crystallin gene in explanted chicken lens epithelia. Proc Natl Acad Sci U S A. 1985 Apr;82(8):2334–2338. doi: 10.1073/pnas.82.8.2334. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chepelinsky A. B., Sommer B., Piatigorsky J. Interaction between two different regulatory elements activates the murine alpha A-crystallin gene promoter in explanted lens epithelia. Mol Cell Biol. 1987 May;7(5):1807–1814. doi: 10.1128/mcb.7.5.1807. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Clerc R. G., Corcoran L. M., LeBowitz J. H., Baltimore D., Sharp P. A. The B-cell-specific Oct-2 protein contains POU box- and homeo box-type domains. Genes Dev. 1988 Dec;2(12A):1570–1581. doi: 10.1101/gad.2.12a.1570. [DOI] [PubMed] [Google Scholar]
  10. Crouse G. F., Frischauf A., Lehrach H. An integrated and simplified approach to cloning into plasmids and single-stranded phages. Methods Enzymol. 1983;101:78–89. doi: 10.1016/0076-6879(83)01006-x. [DOI] [PubMed] [Google Scholar]
  11. Das G. C., Piatigorsky J. The chicken delta 1-crystallin gene promoter: binding of transcription factor(s) to the upstream G+C-rich region is necessary for promoter function in vitro. Proc Natl Acad Sci U S A. 1986 May;83(10):3131–3135. doi: 10.1073/pnas.83.10.3131. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Davidson E. H., Jacobs H. T., Britten R. J. Very short repeats and coordinate induction of genes. Nature. 1983 Feb 10;301(5900):468–470. doi: 10.1038/301468a0. [DOI] [PubMed] [Google Scholar]
  13. Delaye M., Tardieu A. Short-range order of crystallin proteins accounts for eye lens transparency. 1983 Mar 31-Apr 6Nature. 302(5907):415–417. doi: 10.1038/302415a0. [DOI] [PubMed] [Google Scholar]
  14. Emerson B. M., Nickol J. M., Jackson P. D., Felsenfeld G. Analysis of the tissue-specific enhancer at the 3' end of the chicken adult beta-globin gene. Proc Natl Acad Sci U S A. 1987 Jul;84(14):4786–4790. doi: 10.1073/pnas.84.14.4786. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Evans R. M. The steroid and thyroid hormone receptor superfamily. Science. 1988 May 13;240(4854):889–895. doi: 10.1126/science.3283939. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Fernald R. D., Wright S. E. Maintenance of optical quality during crystalline lens growth. Nature. 1983 Feb 17;301(5901):618–620. doi: 10.1038/301618a0. [DOI] [PubMed] [Google Scholar]
  17. Gillies S. D., Morrison S. L., Oi V. T., Tonegawa S. A tissue-specific transcription enhancer element is located in the major intron of a rearranged immunoglobulin heavy chain gene. Cell. 1983 Jul;33(3):717–728. doi: 10.1016/0092-8674(83)90014-4. [DOI] [PubMed] [Google Scholar]
  18. Goodbourn S., Maniatis T. Overlapping positive and negative regulatory domains of the human beta-interferon gene. Proc Natl Acad Sci U S A. 1988 Mar;85(5):1447–1451. doi: 10.1073/pnas.85.5.1447. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Gorman C. M., Moffat L. F., Howard B. H. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol. 1982 Sep;2(9):1044–1051. doi: 10.1128/mcb.2.9.1044. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hall C. V., Jacob P. E., Ringold G. M., Lee F. Expression and regulation of Escherichia coli lacZ gene fusions in mammalian cells. J Mol Appl Genet. 1983;2(1):101–109. [PubMed] [Google Scholar]
  21. Harding J. J., Dilley K. J. Structural proteins of the mammalian lens: a review with emphasis on changes in development, aging and cataract. Exp Eye Res. 1976 Jan;22(1):1–73. doi: 10.1016/0014-4835(76)90033-6. [DOI] [PubMed] [Google Scholar]
  22. Hayashi S., Goto K., Okada T. S., Kondoh H. Lens-specific enhancer in the third intron regulates expression of the chicken delta 1-crystallin gene. Genes Dev. 1987 Oct;1(8):818–828. doi: 10.1101/gad.1.8.818. [DOI] [PubMed] [Google Scholar]
  23. Hayashi S., Kondoh H. In vivo competition of delta-crystallin gene expression by DNA fragments containing a GC box. Mol Cell Biol. 1986 Nov;6(11):4130–4132. doi: 10.1128/mcb.6.11.4130. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Hayashi S., Kondoh H., Yasuda K., Soma G., Ikawa Y., Okada T. S. Tissue-specific regulation of a chicken delta-crystallin gene in mouse cells: involvement of the 5' end region. EMBO J. 1985 Sep;4(9):2201–2207. doi: 10.1002/j.1460-2075.1985.tb03915.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Hejtmancik J. F., Beebe D. C., Ostrer H., Piatigorsky J. delta- and beta-Crystallin mRNA levels in the embryonic and posthatched chicken lens: temporal and spatial changes during development. Dev Biol. 1985 May;109(1):72–81. doi: 10.1016/0012-1606(85)90347-1. [DOI] [PubMed] [Google Scholar]
  26. Hogg D., Tsui L. C., Gorin M., Breitman M. L. Characterization of the human beta-crystallin gene Hu beta A3/A1 reveals ancestral relationships among the beta gamma-crystallin superfamily. J Biol Chem. 1986 Sep 15;261(26):12420–12427. [PubMed] [Google Scholar]
  27. Hope I. A., Struhl K. GCN4, a eukaryotic transcriptional activator protein, binds as a dimer to target DNA. EMBO J. 1987 Sep;6(9):2781–2784. doi: 10.1002/j.1460-2075.1987.tb02573.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Inana G., Piatigorsky J., Norman B., Slingsby C., Blundell T. Gene and protein structure of a beta-crystallin polypeptide in murine lens: relationship of exons and structural motifs. Nature. 1983 Mar 24;302(5906):310–315. doi: 10.1038/302310a0. [DOI] [PubMed] [Google Scholar]
  29. Kondoh H., Yasuda K., Okada T. S. Tissue-specific expression of a cloned chick delta-crystallin gene in mouse cells. Nature. 1983 Feb 3;301(5899):440–442. doi: 10.1038/301440a0. [DOI] [PubMed] [Google Scholar]
  30. Korneluk R. G., Quan F., Gravel R. A. Rapid and reliable dideoxy sequencing of double-stranded DNA. Gene. 1985;40(2-3):317–323. doi: 10.1016/0378-1119(85)90055-1. [DOI] [PubMed] [Google Scholar]
  31. LEABACK D. H., WALKER P. G. Studies on glucosaminidase. 4. The fluorimetric assay of N-acetyl-beta-glucosaminidase. Biochem J. 1961 Jan;78:151–156. doi: 10.1042/bj0780151. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Lenardo M., Pierce J. W., Baltimore D. Protein-binding sites in Ig gene enhancers determine transcriptional activity and inducibility. Science. 1987 Jun 19;236(4808):1573–1577. doi: 10.1126/science.3109035. [DOI] [PubMed] [Google Scholar]
  33. Levy D. E., Kessler D. S., Pine R., Reich N., Darnell J. E., Jr Interferon-induced nuclear factors that bind a shared promoter element correlate with positive and negative transcriptional control. Genes Dev. 1988 Apr;2(4):383–393. doi: 10.1101/gad.2.4.383. [DOI] [PubMed] [Google Scholar]
  34. Lok S., Breitman M. L., Chepelinsky A. B., Piatigorsky J., Gold R. J., Tsui L. C. Lens-specific promoter activity of a mouse gamma-crystallin gene. Mol Cell Biol. 1985 Sep;5(9):2221–2230. doi: 10.1128/mcb.5.9.2221. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Lok S., Tsui L. C., Shinohara T., Piatigorsky J., Gold R., Breitman M. Analysis of the mouse gamma-crystallin gene family: assignment of multiple cDNAs to discrete genomic sequences and characterization of a representative gene. Nucleic Acids Res. 1984 Jun 11;12(11):4517–4529. doi: 10.1093/nar/12.11.4517. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. 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]
  37. McAvoy J. W. Cell division, cell elongation and distribution of alpha-, beta- and gamma-crystallins in the rat lens. J Embryol Exp Morphol. 1978 Apr;44:149–165. [PubMed] [Google Scholar]
  38. McDevitt D. S., Croft L. R. On the existence of gamma-crystallin in the bird lens. Exp Eye Res. 1977 Nov;25(5):473–481. doi: 10.1016/0014-4835(77)90176-2. [DOI] [PubMed] [Google Scholar]
  39. Meakin S. O., Breitman M. L., Tsui L. C. Structural and evolutionary relationships among five members of the human gamma-crystallin gene family. Mol Cell Biol. 1985 Jun;5(6):1408–1414. doi: 10.1128/mcb.5.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Meakin S. O., Du R. P., Tsui L. C., Breitman M. L. Gamma-crystallins of the human eye lens: expression analysis of five members of the gene family. Mol Cell Biol. 1987 Aug;7(8):2671–2679. doi: 10.1128/mcb.7.8.2671. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Melgar E., Goldthwait D. A. Deoxyribonucleic acid nucleases. II. The effects of metals on the mechanism of action of deoxyribonuclease I. J Biol Chem. 1968 Sep 10;243(17):4409–4416. [PubMed] [Google Scholar]
  42. Murer-Orlando M., Paterson R. C., Lok S., Tsui L. C., Breitman M. L. Differential regulation of gamma-crystallin genes during mouse lens development. Dev Biol. 1987 Jan;119(1):260–267. doi: 10.1016/0012-1606(87)90227-2. [DOI] [PubMed] [Google Scholar]
  43. Nir U., Walker M. D., Rutter W. J. Regulation of rat insulin 1 gene expression: evidence for negative regulation in nonpancreatic cells. Proc Natl Acad Sci U S A. 1986 May;83(10):3180–3184. doi: 10.1073/pnas.83.10.3180. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Nomiyama H., Fromental C., Xiao J. H., Chambon P. Cell-specific activity of the constituent elements of the simian virus 40 enhancer. Proc Natl Acad Sci U S A. 1987 Nov;84(22):7881–7885. doi: 10.1073/pnas.84.22.7881. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Okazaki K., Yasuda K., Kondoh H., Okada T. S. DNA sequences responsible for tissue-specific expression of a chicken alpha-crystallin gene in mouse lens cells. EMBO J. 1985 Oct;4(10):2589–2595. doi: 10.1002/j.1460-2075.1985.tb03975.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Ostrer H., Beebe D. C., Piatigorsky J. Beta-crystallin mRNAs: differential distribution in the developing chicken lens. Dev Biol. 1981 Sep;86(2):403–408. doi: 10.1016/0012-1606(81)90198-6. [DOI] [PubMed] [Google Scholar]
  47. Pelham H. R. A regulatory upstream promoter element in the Drosophila hsp 70 heat-shock gene. Cell. 1982 Sep;30(2):517–528. doi: 10.1016/0092-8674(82)90249-5. [DOI] [PubMed] [Google Scholar]
  48. Poellinger L., Yoza B. K., Roeder R. G. Functional cooperativity between protein molecules bound at two distinct sequence elements of the immunoglobulin heavy-chain promoter. Nature. 1989 Feb 9;337(6207):573–576. doi: 10.1038/337573a0. [DOI] [PubMed] [Google Scholar]
  49. Pruijn G. J., van Driel W., van Miltenburg R. T., van der Vliet P. C. Promoter and enhancer elements containing a conserved sequence motif are recognized by nuclear factor III, a protein stimulating adenovirus DNA replication. EMBO J. 1987 Dec 1;6(12):3771–3778. doi: 10.1002/j.1460-2075.1987.tb02712.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Quinlan P., Oda S., Breitman M. L., Tsui L. C. The mouse eye lens obsolescence (Elo) mutant: studies on crystallin gene expression and linkage analysis between the mutant locus and the gamma-crystallin genes. Genes Dev. 1987 Sep;1(7):637–644. doi: 10.1101/gad.1.7.637. [DOI] [PubMed] [Google Scholar]
  51. Remmers E. F., Yang J. Q., Marcu K. B. A negative transcriptional control element located upstream of the murine c-myc gene. EMBO J. 1986 May;5(5):899–904. doi: 10.1002/j.1460-2075.1986.tb04301.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. 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]
  53. Schöler H. R., Gruss P. Specific interaction between enhancer-containing molecules and cellular components. Cell. 1984 Feb;36(2):403–411. doi: 10.1016/0092-8674(84)90233-2. [DOI] [PubMed] [Google Scholar]
  54. Searle P. F., Stuart G. W., Palmiter R. D. Building a metal-responsive promoter with synthetic regulatory elements. Mol Cell Biol. 1985 Jun;5(6):1480–1489. doi: 10.1128/mcb.5.6.1480. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Shuey D. J., Parker C. S. Binding of Drosophila heat-shock gene transcription factor to the hsp 70 promoter. Evidence for symmetric and dynamic interactions. J Biol Chem. 1986 Jun 15;261(17):7934–7940. [PubMed] [Google Scholar]
  56. Siezen R. J., Wu E., Kaplan E. D., Thomson J. A., Benedek G. B. Rat lens gamma-crystallins. Characterization of the six gene products and their spatial and temporal distribution resulting from differential synthesis. J Mol Biol. 1988 Feb 5;199(3):475–490. doi: 10.1016/0022-2836(88)90619-5. [DOI] [PubMed] [Google Scholar]
  57. Sive H. L., Roeder R. G. Interaction of a common factor with conserved promoter and enhancer sequences in histone H2B, immunoglobulin, and U2 small nuclear RNA (snRNA) genes. Proc Natl Acad Sci U S A. 1986 Sep;83(17):6382–6386. doi: 10.1073/pnas.83.17.6382. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Skow L. C., Adkison L., Womack J. E., Beamer W. G., Taylor B. A. Mapping of the mouse fibronectin gene (Fn-1) to chromosome 1: conservation of the Idh-1-Cryg-Fn-1 synteny group in mammals. Genomics. 1987 Nov;1(3):283–286. doi: 10.1016/0888-7543(87)90057-7. [DOI] [PubMed] [Google Scholar]
  59. Stuart G. W., Searle P. F., Palmiter R. D. Identification of multiple metal regulatory elements in mouse metallothionein-I promoter by assaying synthetic sequences. 1985 Oct 31-Nov 6Nature. 317(6040):828–831. doi: 10.1038/317828a0. [DOI] [PubMed] [Google Scholar]
  60. Sturm R. A., Das G., Herr W. The ubiquitous octamer-binding protein Oct-1 contains a POU domain with a homeo box subdomain. Genes Dev. 1988 Dec;2(12A):1582–1599. doi: 10.1101/gad.2.12a.1582. [DOI] [PubMed] [Google Scholar]
  61. Takeda Y., Ohlendorf D. H., Anderson W. F., Matthews B. W. DNA-binding proteins. Science. 1983 Sep 9;221(4615):1020–1026. doi: 10.1126/science.6308768. [DOI] [PubMed] [Google Scholar]
  62. Thompson M. A., Hawkins J. W., Piatigorsky J. Complete nucleotide sequence of the chicken alpha A-crystallin gene and its 5' flanking region. Gene. 1987;56(2-3):173–184. doi: 10.1016/0378-1119(87)90135-1. [DOI] [PubMed] [Google Scholar]
  63. Walker M. D., Edlund T., Boulet A. M., Rutter W. J. Cell-specific expression controlled by the 5'-flanking region of insulin and chymotrypsin genes. Nature. 1983 Dec 8;306(5943):557–561. doi: 10.1038/306557a0. [DOI] [PubMed] [Google Scholar]
  64. Weiher H., König M., Gruss P. Multiple point mutations affecting the simian virus 40 enhancer. Science. 1983 Feb 11;219(4585):626–631. doi: 10.1126/science.6297005. [DOI] [PubMed] [Google Scholar]
  65. Widen S. G., Papaconstantinou J. Extinction of alpha-fetoprotein gene expression in somatic cell hybrids involves cis-acting DNA elements. Mol Cell Biol. 1987 Jul;7(7):2606–2609. doi: 10.1128/mcb.7.7.2606. [DOI] [PMC free article] [PubMed] [Google Scholar]
  66. Wirth T., Staudt L., Baltimore D. An octamer oligonucleotide upstream of a TATA motif is sufficient for lymphoid-specific promoter activity. Nature. 1987 Sep 10;329(6135):174–178. doi: 10.1038/329174a0. [DOI] [PubMed] [Google Scholar]
  67. Wistow G., Slingsby C., Blundell T., Driessen H., De Jong W., Bloemendal H. Eye-lens proteins: the three-dimensional structure of beta-crystallin predicted from monomeric gamma-crystallin. FEBS Lett. 1981 Oct 12;133(1):9–16. doi: 10.1016/0014-5793(81)80460-7. [DOI] [PubMed] [Google Scholar]
  68. Wistow G., Turnell B., Summers L., Slingsby C., Moss D., Miller L., Lindley P., Blundell T. X-ray analysis of the eye lens protein gamma-II crystallin at 1.9 A resolution. J Mol Biol. 1983 Oct 15;170(1):175–202. doi: 10.1016/s0022-2836(83)80232-0. [DOI] [PubMed] [Google Scholar]
  69. den Dunnen J. T., Moormann R. J., Cremers F. P., Schoenmakers J. G. Two human gamma-crystallin genes are linked and riddled with Alu-repeats. Gene. 1985;38(1-3):197–204. doi: 10.1016/0378-1119(85)90218-5. [DOI] [PubMed] [Google Scholar]
  70. den Dunnen J. T., Moormann R. J., Lubsen N. H., Schoenmakers J. G. Concerted and divergent evolution within the rat gamma-crystallin gene family. J Mol Biol. 1986 May 5;189(1):37–46. doi: 10.1016/0022-2836(86)90379-7. [DOI] [PubMed] [Google Scholar]
  71. den Dunnen J. T., Moormann R. J., Lubsen N. H., Schoenmakers J. G. Intron insertions and deletions in the beta/gamma-crystallin gene family: the rat beta B1 gene. Proc Natl Acad Sci U S A. 1986 May;83(9):2855–2859. doi: 10.1073/pnas.83.9.2855. [DOI] [PMC free article] [PubMed] [Google Scholar]
  72. van Leen R. W., van Roozendaal K. E., Lubsen N. H., Schoenmakers J. G. Differential expression of crystallin genes during development of the rat eye lens. Dev Biol. 1987 Apr;120(2):457–464. doi: 10.1016/0012-1606(87)90249-1. [DOI] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES