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. 1994 Apr 11;22(7):1281–1286. doi: 10.1093/nar/22.7.1281

Identification of a lens-specific regulatory region (LSR) of the murine alpha B-crystallin gene.

R Gopal-Srivastava 1, J Piatigorsky 1
PMCID: PMC523654  PMID: 8165144

Abstract

Previous studies have shown that the -661/+44 sequence of the murine alpha B-crystallin gene contains a muscle-preferred enhancer (-426/-257) and can drive the bacterial chloramphenicol acetyltransferase (CAT) gene in the lens, skeletal muscle and heart of transgenic mice. Here we show that transgenic mice carrying a truncated -164/+44 fragment of the alpha B-crystallin gene fused to the CAT gene expressed exclusively in the lens; by contrast mice carrying a -426/+44 fragment of the alpha B gene fused to CAT expressed highly in the lens, skeletal muscle and heart, and slightly in the lung, brain, kidney, spleen and liver. DNase I protection experiments indicated that the -147/-118 sequence is protected by nuclear proteins from alpha TN4-1 lens cell line, but not by nuclear proteins from myotubes of the C2C12 cell line. Site directed mutagenesis of this sequence decreased promoter activity in transiently-transfected lens cells, consistent with this sequence being a lens-specific regulatory region (LSR). We conclude that the -426/-257 enhancer is required for expression in skeletal muscle, heart and possibly other tissues, and that the -164/+44 sequence of the alpha B-crystallin gene is sufficient for expression in the lens of transgenic mice.

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Selected References

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  1. Aoyama A., Fröhli E., Schäfer R., Klemenz R. Alpha B-crystallin expression in mouse NIH 3T3 fibroblasts: glucocorticoid responsiveness and involvement in thermal protection. Mol Cell Biol. 1993 Mar;13(3):1824–1835. doi: 10.1128/mcb.13.3.1824. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bhat S. P., Nagineni C. N. alpha B subunit of lens-specific protein alpha-crystallin is present in other ocular and non-ocular tissues. Biochem Biophys Res Commun. 1989 Jan 16;158(1):319–325. doi: 10.1016/s0006-291x(89)80215-3. [DOI] [PubMed] [Google Scholar]
  3. Blau H. M., Chiu C. P., Webster C. Cytoplasmic activation of human nuclear genes in stable heterocaryons. Cell. 1983 Apr;32(4):1171–1180. doi: 10.1016/0092-8674(83)90300-8. [DOI] [PubMed] [Google Scholar]
  4. Bloemendal H. Lens proteins. CRC Crit Rev Biochem. 1982;12(1):1–38. doi: 10.3109/10409238209105849. [DOI] [PubMed] [Google Scholar]
  5. Bloemendal H., de Jong W. W. Lens proteins and their genes. Prog Nucleic Acid Res Mol Biol. 1991;41:259–281. doi: 10.1016/s0079-6603(08)60012-4. [DOI] [PubMed] [Google Scholar]
  6. Brakenhoff R. H., Guerts van Kessel A. H., Oldenburg M., Wijnen J. T., Bloemendal H., Meera Khan P., Schoenmakers J. G. Human alpha B-crystallin (CRYA2) gene mapped to chromosome 11q12-q23. Hum Genet. 1990 Jul;85(2):237–240. doi: 10.1007/BF00193203. [DOI] [PubMed] [Google Scholar]
  7. Dasgupta S., Hohman T. C., Carper D. Hypertonic stress induces alpha B-crystallin expression. Exp Eye Res. 1992 Mar;54(3):461–470. doi: 10.1016/0014-4835(92)90058-z. [DOI] [PubMed] [Google Scholar]
  8. Dignam J. D., Lebovitz R. M., Roeder R. G. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Res. 1983 Mar 11;11(5):1475–1489. doi: 10.1093/nar/11.5.1475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dubin R. A., Gopal-Srivastava R., Wawrousek E. F., Piatigorsky J. Expression of the murine alpha B-crystallin gene in lens and skeletal muscle: identification of a muscle-preferred enhancer. Mol Cell Biol. 1991 Sep;11(9):4340–4349. doi: 10.1128/mcb.11.9.4340. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dubin R. A., Wawrousek E. F., Piatigorsky J. Expression of the murine alpha B-crystallin gene is not restricted to the lens. Mol Cell Biol. 1989 Mar;9(3):1083–1091. doi: 10.1128/mcb.9.3.1083. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gopal-Srivastava R., Piatigorsky J. The murine alpha B-crystallin/small heat shock protein enhancer: identification of alpha BE-1, alpha BE-2, alpha BE-3, and MRF control elements. Mol Cell Biol. 1993 Nov;13(11):7144–7152. doi: 10.1128/mcb.13.11.7144. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gordon J. W., Scangos G. A., Plotkin D. J., Barbosa J. A., Ruddle F. H. Genetic transformation of mouse embryos by microinjection of purified DNA. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7380–7384. doi: 10.1073/pnas.77.12.7380. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hawkins J. W., Van Keuren M. L., Piatigorsky J., Law M. L., Patterson D., Kao F. T. Confirmation of assignment of the human alpha 1-crystallin gene (CRYA1) to chromosome 21 with regional localization to q22.3. Hum Genet. 1987 Aug;76(4):375–380. doi: 10.1007/BF00272448. [DOI] [PubMed] [Google Scholar]
  14. Iwaki T., Kume-Iwaki A., Liem R. K., Goldman J. E. Alpha B-crystallin is expressed in non-lenticular tissues and accumulates in Alexander's disease brain. Cell. 1989 Apr 7;57(1):71–78. doi: 10.1016/0092-8674(89)90173-6. [DOI] [PubMed] [Google Scholar]
  15. Kato K., Shinohara H., Kurobe N., Goto S., Inaguma Y., Ohshima K. Immunoreactive alpha A crystallin in rat non-lenticular tissues detected with a sensitive immunoassay method. Biochim Biophys Acta. 1991 Oct 25;1080(2):173–180. doi: 10.1016/0167-4838(91)90146-q. [DOI] [PubMed] [Google Scholar]
  16. Klemenz R., Fröhli E., Aoyama A., Hoffmann S., Simpson R. J., Moritz R. L., Schäfer R. Alpha B crystallin accumulation is a specific response to Ha-ras and v-mos oncogene expression in mouse NIH 3T3 fibroblasts. Mol Cell Biol. 1991 Feb;11(2):803–812. doi: 10.1128/mcb.11.2.803. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Klemenz R., Fröhli E., Steiger R. H., Schäfer R., Aoyama A. Alpha B-crystallin is a small heat shock protein. Proc Natl Acad Sci U S A. 1991 May 1;88(9):3652–3656. doi: 10.1073/pnas.88.9.3652. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Klemenz R., Hoffmann S., Jaggi R., Werenskiold A. K. The v-mos and c-Ha-ras oncoproteins exert similar effects on the pattern of protein synthesis. Oncogene. 1989 Jun;4(6):799–803. [PubMed] [Google Scholar]
  19. Lowe J., McDermott H., Pike I., Spendlove I., Landon M., Mayer R. J. alpha B crystallin expression in non-lenticular tissues and selective presence in ubiquitinated inclusion bodies in human disease. J Pathol. 1992 Jan;166(1):61–68. doi: 10.1002/path.1711660110. [DOI] [PubMed] [Google Scholar]
  20. Matsuo I., Takeuchi M., Yasuda K. Identification of the contact sites of a factor that interacts with motif I (alpha CE1) of the chicken alpha A-crystallin lens-specific enhancer. Biochem Biophys Res Commun. 1992 Apr 15;184(1):24–30. doi: 10.1016/0006-291x(92)91152-g. [DOI] [PubMed] [Google Scholar]
  21. Matsuo I., Yasuda K. The cooperative interaction between two motifs of an enhancer element of the chicken alpha A-crystallin gene, alpha CE1 and alpha CE2, confers lens-specific expression. Nucleic Acids Res. 1992 Jul 25;20(14):3701–3712. doi: 10.1093/nar/20.14.3701. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Ngo J. T., Klisak I., Dubin R. A., Piatigorsky J., Mohandas T., Sparkes R. S., Bateman J. B. Assignment of the alpha B-crystallin gene to human chromosome 11. Genomics. 1989 Nov;5(4):665–669. doi: 10.1016/0888-7543(89)90106-7. [DOI] [PubMed] [Google Scholar]
  23. Piatigorsky J. Lens crystallins. Innovation associated with changes in gene regulation. J Biol Chem. 1992 Mar 5;267(7):4277–4280. [PubMed] [Google Scholar]
  24. Piatigorsky J., Wistow G. J. Enzyme/crystallins: gene sharing as an evolutionary strategy. Cell. 1989 Apr 21;57(2):197–199. doi: 10.1016/0092-8674(89)90956-2. [DOI] [PubMed] [Google Scholar]
  25. Piatigorsky J., Wistow G. The recruitment of crystallins: new functions precede gene duplication. Science. 1991 May 24;252(5009):1078–1079. doi: 10.1126/science.252.5009.1078. [DOI] [PubMed] [Google Scholar]
  26. Reddan J. R., Chepelinsky A. B., Dziedzic D. C., Piatigorsky J., Goldenberg E. M. Retention of lens specificity in long-term cultures of diploid rabbit lens epithelial cells. Differentiation. 1986;33(2):168–174. doi: 10.1111/j.1432-0436.1986.tb00422.x. [DOI] [PubMed] [Google Scholar]
  27. Srinivasan A. N., Nagineni C. N., Bhat S. P. alpha A-crystallin is expressed in non-ocular tissues. J Biol Chem. 1992 Nov 15;267(32):23337–23341. [PubMed] [Google Scholar]
  28. Wistow G. J., Piatigorsky J. Lens crystallins: the evolution and expression of proteins for a highly specialized tissue. Annu Rev Biochem. 1988;57:479–504. doi: 10.1146/annurev.bi.57.070188.002403. [DOI] [PubMed] [Google Scholar]
  29. Yamada T., Nakamura T., Westphal H., Russell P. Synthesis of alpha-crystallin by a cell line derived from the lens of a transgenic animal. Curr Eye Res. 1990 Jan;9(1):31–37. doi: 10.3109/02713689009000052. [DOI] [PubMed] [Google Scholar]
  30. de Jong W. W., Hendriks W., Mulders J. W., Bloemendal H. Evolution of eye lens crystallins: the stress connection. Trends Biochem Sci. 1989 Sep;14(9):365–368. doi: 10.1016/0968-0004(89)90009-1. [DOI] [PubMed] [Google Scholar]
  31. de Jong W. W., Leunissen J. A., Voorter C. E. Evolution of the alpha-crystallin/small heat-shock protein family. Mol Biol Evol. 1993 Jan;10(1):103–126. doi: 10.1093/oxfordjournals.molbev.a039992. [DOI] [PubMed] [Google Scholar]

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