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. 1991 Feb;10(2):255–261. doi: 10.1002/j.1460-2075.1991.tb07945.x

The murine Spi-2 proteinase inhibitor locus: a multigene family with a hypervariable reactive site domain.

J D Inglis 1, R E Hill 1
PMCID: PMC452641  PMID: 1991447

Abstract

We have isolated 10 closely linked members of a proteinase inhibitor multigene family from the inbred mouse strain 129. These sequences, termed the Serine Proteinase Inhibitor 2 (Spi-2) genes, appear to have been derived from a common ancestor represented in man by the single copy alpha 1-antichymotrypsin gene. The genes are clustered on two cloned genomic DNA segments spanning 220 kb, and have at least partially retained the intragenic structure of the ancestral Spi-2 gene. Sequence analysis from the final coding exon indicates that most of the mouse genes may be competent to encode functional proteins, some with a predictable inhibitory spectrum, and several representing novel inhibitor types. An oligonucleotide probe designed to one reactive centre sequence enabled the isolation of the cognate expressed transcript from a liver cDNA library. However, whether expressed or not, the reactive centre regions of all the sequences have diverged at a rapid rate relative to structurally defined flanking sequences. The divergence is also appreciably greater than that occurring in an adjacent non-coding sequence. This phenomenon has established novel potential inhibitory specificities, while maintaining a functional inhibitor structure.

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Selected References

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

  1. Bennett K. L., Hastie N. D. Looking for relationships between the most repeated dispersed DNA sequences in the mouse: small R elements are found associated consistently with long MIF repeats. EMBO J. 1984 Feb;3(2):467–472. doi: 10.1002/j.1460-2075.1984.tb01829.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bennett K. L., Hill R. E., Pietras D. F., Woodworth-Gutai M., Kane-Haas C., Houston J. M., Heath J. K., Hastie N. D. Most highly repeated dispersed DNA families in the mouse genome. Mol Cell Biol. 1984 Aug;4(8):1561–1571. doi: 10.1128/mcb.4.8.1561. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bentley D. L., Rabbitts T. H. Evolution of immunoglobulin V genes: evidence indicating that recently duplicated human V kappa sequences have diverged by gene conversion. Cell. 1983 Jan;32(1):181–189. doi: 10.1016/0092-8674(83)90508-1. [DOI] [PubMed] [Google Scholar]
  4. Bird A. P. CpG-rich islands and the function of DNA methylation. Nature. 1986 May 15;321(6067):209–213. doi: 10.1038/321209a0. [DOI] [PubMed] [Google Scholar]
  5. Bode W., Papamokos E., Musil D., Seemueller U., Fritz H. Refined 1.2 A crystal structure of the complex formed between subtilisin Carlsberg and the inhibitor eglin c. Molecular structure of eglin and its detailed interaction with subtilisin. EMBO J. 1986 Apr;5(4):813–818. doi: 10.1002/j.1460-2075.1986.tb04286.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bode W., Wei A. Z., Huber R., Meyer E., Travis J., Neumann S. X-ray crystal structure of the complex of human leukocyte elastase (PMN elastase) and the third domain of the turkey ovomucoid inhibitor. EMBO J. 1986 Oct;5(10):2453–2458. doi: 10.1002/j.1460-2075.1986.tb04521.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Carrell R. W., Aulak K. S., Owen M. C. The molecular pathology of the serpins. Mol Biol Med. 1989 Feb;6(1):35–42. [PubMed] [Google Scholar]
  8. Carrell R. W., Pemberton P. A., Boswell D. R. The serpins: evolution and adaptation in a family of protease inhibitors. Cold Spring Harb Symp Quant Biol. 1987;52:527–535. doi: 10.1101/sqb.1987.052.01.060. [DOI] [PubMed] [Google Scholar]
  9. Chao J., Chai K. X., Chen L. M., Xiong W., Chao S., Woodley-Miller C., Wang L. X., Lu H. S., Chao L. Tissue kallikrein-binding protein is a serpin. I. Purification, characterization, and distribution in normotensive and spontaneously hypertensive rats. J Biol Chem. 1990 Sep 25;265(27):16394–16401. [PubMed] [Google Scholar]
  10. Clark A. J., Chave-Cox A., Ma X., Bishop J. O. Analysis of mouse major urinary protein genes: variation between the exonic sequences of group 1 genes and a comparison with an active gene out with group 1 both suggest that gene conversion has occurred between MUP genes. EMBO J. 1985 Dec 1;4(12):3167–3171. doi: 10.1002/j.1460-2075.1985.tb04060.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Courtney M., Jallat S., Tessier L. H., Benavente A., Crystal R. G., Lecocq J. P. Synthesis in E. coli of alpha 1-antitrypsin variants of therapeutic potential for emphysema and thrombosis. Nature. 1985 Jan 10;313(5998):149–151. doi: 10.1038/313149a0. [DOI] [PubMed] [Google Scholar]
  12. Creighton T. E., Darby N. J. Functional evolutionary divergence of proteolytic enzymes and their inhibitors. Trends Biochem Sci. 1989 Aug;14(8):319–324. doi: 10.1016/0968-0004(89)90159-x. [DOI] [PubMed] [Google Scholar]
  13. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ehrich E., Craig A., Poustka A., Frischauf A. M., Lehrach H. A family of cosmid vectors with the multi-copy R6K replication origin. Gene. 1987;57(2-3):229–237. doi: 10.1016/0378-1119(87)90126-0. [DOI] [PubMed] [Google Scholar]
  15. Evans B. A., Drinkwater C. C., Richards R. I. Mouse glandular kallikrein genes. Structure and partial sequence analysis of the kallikrein gene locus. J Biol Chem. 1987 Jun 15;262(17):8027–8034. [PubMed] [Google Scholar]
  16. Flink I. L., Bailey T. J., Gustafson T. A., Markham B. E., Morkin E. Complete amino acid sequence of human thyroxine-binding globulin deduced from cloned DNA: close homology to the serine antiproteases. Proc Natl Acad Sci U S A. 1986 Oct;83(20):7708–7712. doi: 10.1073/pnas.83.20.7708. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hammond G. L., Smith C. L., Goping I. S., Underhill D. A., Harley M. J., Reventos J., Musto N. A., Gunsalus G. L., Bardin C. W. Primary structure of human corticosteroid binding globulin, deduced from hepatic and pulmonary cDNAs, exhibits homology with serine protease inhibitors. Proc Natl Acad Sci U S A. 1987 Aug;84(15):5153–5157. doi: 10.1073/pnas.84.15.5153. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hill R. E., Hastie N. D. Accelerated evolution in the reactive centre regions of serine protease inhibitors. Nature. 1987 Mar 5;326(6108):96–99. doi: 10.1038/326096a0. [DOI] [PubMed] [Google Scholar]
  19. Hill R. E., Shaw P. H., Barth R. K., Hastie N. D. A genetic locus closely linked to a protease inhibitor gene complex controls the level of multiple RNA transcripts. Mol Cell Biol. 1985 Aug;5(8):2114–2122. doi: 10.1128/mcb.5.8.2114. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hill R. E., Shaw P. H., Boyd P. A., Baumann H., Hastie N. D. Plasma protease inhibitors in mouse and man: divergence within the reactive centre regions. Nature. 1984 Sep 13;311(5982):175–177. doi: 10.1038/311175a0. [DOI] [PubMed] [Google Scholar]
  21. Holmes W. E., Lijnen H. R., Collen D. Characterization of recombinant human alpha 2-antiplasmin and of mutants obtained by site-directed mutagenesis of the reactive site. Biochemistry. 1987 Aug 11;26(16):5133–5140. doi: 10.1021/bi00390a036. [DOI] [PubMed] [Google Scholar]
  22. Huber R., Carrell R. W. Implications of the three-dimensional structure of alpha 1-antitrypsin for structure and function of serpins. Biochemistry. 1989 Nov 14;28(23):8951–8966. doi: 10.1021/bi00449a001. [DOI] [PubMed] [Google Scholar]
  23. Hunt L. T., Dayhoff M. O. A surprising new protein superfamily containing ovalbumin, antithrombin-III, and alpha 1-proteinase inhibitor. Biochem Biophys Res Commun. 1980 Jul 31;95(2):864–871. doi: 10.1016/0006-291x(80)90867-0. [DOI] [PubMed] [Google Scholar]
  24. Jallat S., Carvallo D., Tessier L. H., Roecklin D., Roitsch C., Ogushi F., Crystal R. G., Courtney M. Altered specificities of genetically engineered alpha 1 antitrypsin variants. Protein Eng. 1986 Oct-Nov;1(1):29–35. doi: 10.1093/protein/1.1.29. [DOI] [PubMed] [Google Scholar]
  25. Laskowski M., Jr, Kato I. Protein inhibitors of proteinases. Annu Rev Biochem. 1980;49:593–626. doi: 10.1146/annurev.bi.49.070180.003113. [DOI] [PubMed] [Google Scholar]
  26. Lawlor D. A., Zemmour J., Ennis P. D., Parham P. Evolution of class-I MHC genes and proteins: from natural selection to thymic selection. Annu Rev Immunol. 1990;8:23–63. doi: 10.1146/annurev.iy.08.040190.000323. [DOI] [PubMed] [Google Scholar]
  27. Levy N. J., Ramesh N., Cicardi M., Harrison R. A., Davis A. E., 3rd Type II hereditary angioneurotic edema that may result from a single nucleotide change in the codon for alanine-436 in the C1 inhibitor gene. Proc Natl Acad Sci U S A. 1990 Jan;87(1):265–268. doi: 10.1073/pnas.87.1.265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Loebermann H., Tokuoka R., Deisenhofer J., Huber R. Human alpha 1-proteinase inhibitor. Crystal structure analysis of two crystal modifications, molecular model and preliminary analysis of the implications for function. J Mol Biol. 1984 Aug 15;177(3):531–557. [PubMed] [Google Scholar]
  29. Maeda N., Smithies O. The evolution of multigene families: human haptoglobin genes. Annu Rev Genet. 1986;20:81–108. doi: 10.1146/annurev.ge.20.120186.000501. [DOI] [PubMed] [Google Scholar]
  30. Ohkubo H., Kageyama R., Ujihara M., Hirose T., Inayama S., Nakanishi S. Cloning and sequence analysis of cDNA for rat angiotensinogen. Proc Natl Acad Sci U S A. 1983 Apr;80(8):2196–2200. doi: 10.1073/pnas.80.8.2196. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Owen M. C., Brennan S. O., Lewis J. H., Carrell R. W. Mutation of antitrypsin to antithrombin. alpha 1-antitrypsin Pittsburgh (358 Met leads to Arg), a fatal bleeding disorder. N Engl J Med. 1983 Sep 22;309(12):694–698. doi: 10.1056/NEJM198309223091203. [DOI] [PubMed] [Google Scholar]
  32. Perry D. J., Carrell R. W. CpG dinucleotides are "hotspots" for mutation in the antithrombin III gene. Twelve variants identified using the polymerase chain reaction. Mol Biol Med. 1989 Jun;6(3):239–243. [PubMed] [Google Scholar]
  33. Rogers J. H. The origin and evolution of retroposons. Int Rev Cytol. 1985;93:187–279. doi: 10.1016/s0074-7696(08)61375-3. [DOI] [PubMed] [Google Scholar]
  34. Schaffer H. E., Sederoff R. R. Improved estimation of DNA fragment lengths from Agarose gels. Anal Biochem. 1981 Jul 15;115(1):113–122. doi: 10.1016/0003-2697(81)90533-9. [DOI] [PubMed] [Google Scholar]
  35. Slightom J. L., Blechl A. E., Smithies O. Human fetal G gamma- and A gamma-globin genes: complete nucleotide sequences suggest that DNA can be exchanged between these duplicated genes. Cell. 1980 Oct;21(3):627–638. doi: 10.1016/0092-8674(80)90426-2. [DOI] [PubMed] [Google Scholar]
  36. Stein P. E., Leslie A. G., Finch J. T., Turnell W. G., McLaughlin P. J., Carrell R. W. Crystal structure of ovalbumin as a model for the reactive centre of serpins. Nature. 1990 Sep 6;347(6288):99–102. doi: 10.1038/347099a0. [DOI] [PubMed] [Google Scholar]
  37. Stephens A. W., Siddiqui A., Hirs C. H. Site-directed mutagenesis of the reactive center (serine 394) of antithrombin III. J Biol Chem. 1988 Nov 5;263(31):15849–15852. [PubMed] [Google Scholar]
  38. Suzuki K., Deyashiki Y., Nishioka J., Kurachi K., Akira M., Yamamoto S., Hashimoto S. Characterization of a cDNA for human protein C inhibitor. A new member of the plasma serine protease inhibitor superfamily. J Biol Chem. 1987 Jan 15;262(2):611–616. [PubMed] [Google Scholar]
  39. Takahara H., Sinohara H. Mouse plasma trypsin inhibitors. Isolation and characterization of alpha-1-antitrypsin and contrapsin, a novel trypsin inhibitor. J Biol Chem. 1982 Mar 10;257(5):2438–2446. [PubMed] [Google Scholar]
  40. Travis J., Salvesen G. S. Human plasma proteinase inhibitors. Annu Rev Biochem. 1983;52:655–709. doi: 10.1146/annurev.bi.52.070183.003255. [DOI] [PubMed] [Google Scholar]
  41. Weiss E., Golden L., Zakut R., Mellor A., Fahrner K., Kvist S., Flavell R. A. The DNA sequence of the H-2kb gene: evidence for gene conversion as a mechanism for the generation of polymorphism in histocompatibilty antigens. EMBO J. 1983;2(3):453–462. doi: 10.1002/j.1460-2075.1983.tb01444.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Wright H. T., Qian H. X., Huber R. Crystal structure of plakalbumin, a proteolytically nicked form of ovalbumin. Its relationship to the structure of cleaved alpha-1-proteinase inhibitor. J Mol Biol. 1990 Jun 5;213(3):513–528. doi: 10.1016/s0022-2836(05)80212-8. [DOI] [PubMed] [Google Scholar]

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