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. 1991 Mar 15;88(6):2456–2460. doi: 10.1073/pnas.88.6.2456

Multiple forms of mRNA encoding human pregnancy-associated endometrial alpha 2-globulin, a beta-lactoglobulin homologue.

J Garde 1, S C Bell 1, I C Eperon 1
PMCID: PMC51251  PMID: 2006183

Abstract

Human pregnancy-associated endometrial alpha 2-globulin (alpha 2-PEG) is the major secretory protein product of the endometrium during embryo implantation and the first few weeks of pregnancy. It is a homologue of beta-lactoglobulin, a retinol binding protein, but unlike beta-lactoglobulin it is not found in the mammary gland. The cloning and sequencing of 34 alpha 2-PEG clones has revealed several minor variant forms indicative of alternatively spliced alpha 2-PEG pre-mRNA. These minor forms have also been detected amongst uncloned cDNA after PCR amplification. Some of these mRNAs would give rise to forms of alpha 2-PEG protein lacking internal sequences, whereas others affect the mRNA sequences on the 3' boundary of the presumed termination codon. Sequences within the cDNA clones are consistent with the existence of splice sites, and together with similarities found between alpha 2-PEG cDNA and beta-lactoglobulin gene sequences there is good evidence in support of an unusual scheme for the alternative splicing of alpha 2-PEG pre-mRNA involving both alternative 5' splice sites and alternative 3' splice sites. This scheme suggests that the alpha 2-PEG and beta-lactoglobulin genes share a similar structure in at least two regions, and it is likely that beta-lactoglobulin pre-mRNA would show a similar pattern of alternative splicing for one of these regions.

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Selected References

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  1. Ali S., Clark A. J. Characterization of the gene encoding ovine beta-lactoglobulin. Similarity to the genes for retinol binding protein and other secretory proteins. J Mol Biol. 1988 Feb 5;199(3):415–426. doi: 10.1016/0022-2836(88)90614-6. [DOI] [PubMed] [Google Scholar]
  2. Bell S. C., Doré-Green F. Detection and characterization of human secretory "pregnancy-associated endometrial alpha 2-globulin" in uterine luminal fluid. J Reprod Immunol. 1987 May;11(1):13–29. doi: 10.1016/0165-0378(87)90003-9. [DOI] [PubMed] [Google Scholar]
  3. Bell S. C., Hales M. W., Patel S. R., Kirwan P. H., Drife J. O., Milford-Ward A. Amniotic fluid concentrations of secreted pregnancy-associated endometrial alpha 1- and alpha 2-globulins (alpha 1- and alpha 2-PEG). Br J Obstet Gynaecol. 1986 Sep;93(9):909–915. doi: 10.1111/j.1471-0528.1986.tb08007.x. [DOI] [PubMed] [Google Scholar]
  4. Bell S. C., Hales M. W., Patel S., Kirwan P. H., Drife J. O. Protein synthesis and secretion by the human endometrium and decidua during early pregnancy. Br J Obstet Gynaecol. 1985 Aug;92(8):793–803. doi: 10.1111/j.1471-0528.1985.tb03048.x. [DOI] [PubMed] [Google Scholar]
  5. Bell S. C., Keyte J. W., Waites G. T. Pregnancy-associated endometrial alpha 2-globulin, the major secretory protein of the luteal phase and first trimester pregnancy endometrium, is not glycosylated prolactin but related to beta-lactoglobulins. J Clin Endocrinol Metab. 1987 Nov;65(5):1067–1071. doi: 10.1210/jcem-65-5-1067. [DOI] [PubMed] [Google Scholar]
  6. Bell S. C., Patel S. R., Kirwan P. H., Drife J. O. Protein synthesis and secretion by the human endometrium during the menstrual cycle and the effect of progesterone in vitro. J Reprod Fertil. 1986 May;77(1):221–231. doi: 10.1530/jrf.0.0770221. [DOI] [PubMed] [Google Scholar]
  7. Bell S. C., Patel S. R., Kirwan P. H., Drife J. O. Protein synthesis and secretion by the human endometrium during the menstrual cycle and the effect of progesterone in vitro. J Reprod Fertil. 1986 May;77(1):221–231. doi: 10.1530/jrf.0.0770221. [DOI] [PubMed] [Google Scholar]
  8. Bell S. C. Purification of human secretory pregnancy-associated endometrial alpha 2-globulin (alpha 2-PEG) from cytosol of first trimester pregnancy endometrium. Hum Reprod. 1986 Aug;1(5):313–318. doi: 10.1093/oxfordjournals.humrep.a136412. [DOI] [PubMed] [Google Scholar]
  9. Bell S. C. Secretory endometrial and decidual proteins: studies and clinical significance of a maternally derived group of pregnancy-associated serum proteins. Hum Reprod. 1986 Apr;1(3):129–143. doi: 10.1093/oxfordjournals.humrep.a136369. [DOI] [PubMed] [Google Scholar]
  10. Bernstein P., Ross J. Poly(A), poly(A) binding protein and the regulation of mRNA stability. Trends Biochem Sci. 1989 Sep;14(9):373–377. doi: 10.1016/0968-0004(89)90011-x. [DOI] [PubMed] [Google Scholar]
  11. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  12. Cunningham S. A., Else A. J., Potter B. V., Eperon I. C. Influences of separation and adjacent sequences on the use of alternative 5' splice sites. J Mol Biol. 1991 Jan 20;217(2):265–281. doi: 10.1016/0022-2836(91)90541-d. [DOI] [PubMed] [Google Scholar]
  13. Dobberstein B., Garoff H., Warren G., Robinson P. J. Cell-free synthesis and membrane insertion of mouse H-2Dd histocompatibility antigen and beta 2-microglobulin. Cell. 1979 Aug;17(4):759–769. doi: 10.1016/0092-8674(79)90316-7. [DOI] [PubMed] [Google Scholar]
  14. Eperon L. P., Estibeiro J. P., Eperon I. C. The role of nucleotide sequences in splice site selection in eukaryotic pre-messenger RNA. Nature. 1986 Nov 20;324(6094):280–282. doi: 10.1038/324280a0. [DOI] [PubMed] [Google Scholar]
  15. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  16. Fugate R. D., Song P. S. Spectroscopic characterization of beta-lactoglobulin-retinol complex. Biochim Biophys Acta. 1980 Sep 23;625(1):28–42. doi: 10.1016/0005-2795(80)90105-1. [DOI] [PubMed] [Google Scholar]
  17. Gaye P., Hue-Delahaie D., Mercier J. C., Soulier S., Vilotte J. L., Furet J. P. Ovine beta-lactoglobulin messenger RNA: nucleotide sequence and mRNA levels during functional differentiation of the mammary gland. Biochimie. 1986 Sep;68(9):1097–1107. doi: 10.1016/s0300-9084(86)80184-5. [DOI] [PubMed] [Google Scholar]
  18. Gubler U., Hoffman B. J. A simple and very efficient method for generating cDNA libraries. Gene. 1983 Nov;25(2-3):263–269. doi: 10.1016/0378-1119(83)90230-5. [DOI] [PubMed] [Google Scholar]
  19. Harris S., Ali S., Anderson S., Archibald A. L., Clark A. J. Complete nucleotide sequence of the genomic ovine beta-lactoglobulin gene. Nucleic Acids Res. 1988 Nov 11;16(21):10379–10380. doi: 10.1093/nar/16.21.10379. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Huhtala M. L., Seppälä M., Närvänen A., Palomäki P., Julkunen M., Bohn H. Amino acid sequence homology between human placental protein 14 and beta-lactoglobulins from various species. Endocrinology. 1987 Jun;120(6):2620–2622. doi: 10.1210/endo-120-6-2620. [DOI] [PubMed] [Google Scholar]
  21. Jamieson A. C., Vandeyar M. A., Kang Y. C., Kinsella J. E., Batt C. A. Cloning and nucleotide sequence of the bovine beta-lactoglobulin gene. Gene. 1987;61(1):85–90. doi: 10.1016/0378-1119(87)90367-2. [DOI] [PubMed] [Google Scholar]
  22. Julkunen M., Seppälä M., Jänne O. A. Complete amino acid sequence of human placental protein 14: a progesterone-regulated uterine protein homologous to beta-lactoglobulins. Proc Natl Acad Sci U S A. 1988 Dec;85(23):8845–8849. doi: 10.1073/pnas.85.23.8845. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lear A. L., Eperon L. P., Wheatley I. M., Eperon I. C. Hierarchy for 5' splice site preference determined in vivo. J Mol Biol. 1990 Jan 5;211(1):103–115. doi: 10.1016/0022-2836(90)90014-D. [DOI] [PubMed] [Google Scholar]
  24. Mount S. M. A catalogue of splice junction sequences. Nucleic Acids Res. 1982 Jan 22;10(2):459–472. doi: 10.1093/nar/10.2.459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Nelson K. K., Green M. R. Splice site selection and ribonucleoprotein complex assembly during in vitro pre-mRNA splicing. Genes Dev. 1988 Mar;2(3):319–329. doi: 10.1101/gad.2.3.319. [DOI] [PubMed] [Google Scholar]
  26. Nielsen D. A., Shapiro D. J. Preparation of capped RNA transcripts using T7 RNA polymerase. Nucleic Acids Res. 1986 Jul 25;14(14):5936–5936. doi: 10.1093/nar/14.14.5936. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Noble J. C., Prives C., Manley J. L. Alternative splicing of SV40 early pre-mRNA is determined by branch site selection. Genes Dev. 1988 Nov;2(11):1460–1475. doi: 10.1101/gad.2.11.1460. [DOI] [PubMed] [Google Scholar]
  28. Ohshima Y., Gotoh Y. Signals for the selection of a splice site in pre-mRNA. Computer analysis of splice junction sequences and like sequences. J Mol Biol. 1987 May 20;195(2):247–259. doi: 10.1016/0022-2836(87)90647-4. [DOI] [PubMed] [Google Scholar]
  29. Padgett R. A., Grabowski P. J., Konarska M. M., Seiler S., Sharp P. A. Splicing of messenger RNA precursors. Annu Rev Biochem. 1986;55:1119–1150. doi: 10.1146/annurev.bi.55.070186.005351. [DOI] [PubMed] [Google Scholar]
  30. Papiz M. Z., Sawyer L., Eliopoulos E. E., North A. C., Findlay J. B., Sivaprasadarao R., Jones T. A., Newcomer M. E., Kraulis P. J. The structure of beta-lactoglobulin and its similarity to plasma retinol-binding protein. 1986 Nov 27-Dec 3Nature. 324(6095):383–385. doi: 10.1038/324383a0. [DOI] [PubMed] [Google Scholar]
  31. Reed R., Maniatis T. A role for exon sequences and splice-site proximity in splice-site selection. Cell. 1986 Aug 29;46(5):681–690. doi: 10.1016/0092-8674(86)90343-0. [DOI] [PubMed] [Google Scholar]
  32. Reed R., Maniatis T. The role of the mammalian branchpoint sequence in pre-mRNA splicing. Genes Dev. 1988 Oct;2(10):1268–1276. doi: 10.1101/gad.2.10.1268. [DOI] [PubMed] [Google Scholar]
  33. Reed R. The organization of 3' splice-site sequences in mammalian introns. Genes Dev. 1989 Dec;3(12B):2113–2123. doi: 10.1101/gad.3.12b.2113. [DOI] [PubMed] [Google Scholar]
  34. Robberson B. L., Cote G. J., Berget S. M. Exon definition may facilitate splice site selection in RNAs with multiple exons. Mol Cell Biol. 1990 Jan;10(1):84–94. doi: 10.1128/mcb.10.1.84. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. 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]
  36. Shapiro M. B., Senapathy P. RNA splice junctions of different classes of eukaryotes: sequence statistics and functional implications in gene expression. Nucleic Acids Res. 1987 Sep 11;15(17):7155–7174. doi: 10.1093/nar/15.17.7155. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Shaw G., Kamen R. A conserved AU sequence from the 3' untranslated region of GM-CSF mRNA mediates selective mRNA degradation. Cell. 1986 Aug 29;46(5):659–667. doi: 10.1016/0092-8674(86)90341-7. [DOI] [PubMed] [Google Scholar]
  38. Smith C. W., Porro E. B., Patton J. G., Nadal-Ginard B. Scanning from an independently specified branch point defines the 3' splice site of mammalian introns. Nature. 1989 Nov 16;342(6247):243–247. doi: 10.1038/342243a0. [DOI] [PubMed] [Google Scholar]
  39. Spritz R. A., Strunk K., Surowy C. S., Hoch S. O., Barton D. E., Francke U. The human U1-70K snRNP protein: cDNA cloning, chromosomal localization, expression, alternative splicing and RNA-binding. Nucleic Acids Res. 1987 Dec 23;15(24):10373–10391. doi: 10.1093/nar/15.24.10373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Waites G. T., Bell S. C. Immunohistological localization of human pregnancy-associated endometrial alpha 2-globulin (alpha 2-PEG), a glycosylated beta-lactoglobulin homologue, in the decidua and placenta during pregnancy. J Reprod Fertil. 1989 Sep;87(1):291–300. doi: 10.1530/jrf.0.0870291. [DOI] [PubMed] [Google Scholar]
  41. Waites G. T., Wood P. L., Walker R. A., Bell S. C. Immunohistological localization of human endometrial secretory protein, 'pregnancy-associated endometrial alpha 2-globulin' (alpha 2-PEG), during the menstrual cycle. J Reprod Fertil. 1988 Mar;82(2):665–672. doi: 10.1530/jrf.0.0820665. [DOI] [PubMed] [Google Scholar]
  42. Westwood O. M., Chapman M. G., Totty N., Philp R., Bolton A. E., Lazarus N. R. N-terminal sequence analysis of human placental protein 14, purified in high yield from decidual cytosol. J Reprod Fertil. 1988 Mar;82(2):493–500. doi: 10.1530/jrf.0.0820493. [DOI] [PubMed] [Google Scholar]
  43. Wood P. L., Walker R. A., Bell S. C. Serum levels of pregnancy-associated endometrial alpha 2-globulin (alpha 2-PEG) during normal menstrual and combined oral contraceptive cycles and relationship to immunohistological localization. Hum Reprod. 1989 Feb;4(2):140–146. doi: 10.1093/oxfordjournals.humrep.a136859. [DOI] [PubMed] [Google Scholar]
  44. Zhuang Y. A., Goldstein A. M., Weiner A. M. UACUAAC is the preferred branch site for mammalian mRNA splicing. Proc Natl Acad Sci U S A. 1989 Apr;86(8):2752–2756. doi: 10.1073/pnas.86.8.2752. [DOI] [PMC free article] [PubMed] [Google Scholar]

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