FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / BMC Evol Biol / Alternative splicing after gene duplication drives CEACAM1-p...
BMC evolutionary biology2018; 18(1); 32; doi: 10.1186/s12862-018-1145-x

Alternative splicing after gene duplication drives CEACAM1-paralog diversification in the horse.

Abstract: The CEA gene family is one of the most rapidly evolving gene families in the human genome. The founder gene of the family is thought to be an ancestor of the inhibitory immune checkpoint molecule CEACAM1. Comprehensive analyses of mammalian genomes showed that the CEA gene family is subject to tremendous gene family expansion and contraction events in different mammalian species. While in some species (e.g. rabbits) less than three CEACAM1 related genes exist, were in others (certain microbat species) up to 100 CEACAM1 paralogs identified. We have recently reported that the horse has also an extended CEA gene family. Since mechanisms of gene family expansion and diversification are not well understood we aimed to analyze the equine CEA gene family in detail. We found that the equine CEA gene family contains 17 functional CEACAM1-related genes. Nine of them were secreted molecules and eight CEACAMs contain transmembrane and cytoplasmic domain exons, the latter being in the focus of the present report. Only one (CEACAM41) gene has exons coding for activating signaling motifs all other CEACAM1 paralogs contain cytoplasmic exons similar to that of the inhibitory receptor CEACAM1. However, cloning of cDNAs showed that only one CEACAM1 paralog contain functional immunoreceptor tyrosine-based inhibitory motifs in its cytoplasmic tail. Three receptors have acquired a stop codon in the transmembrane domain and two have lost their inhibitory motifs due to alternative splicing events. In addition, alternative splicing eliminated the transmembrane exon sequence of the putative activating receptor, rendering it to a secreted molecule. Transfection of eukaryotic cells with FLAG-tagged alternatively spliced CEACAMs indicates that they can be expressed in vivo. Thus detection of CEACAM41 mRNA in activated PBMC suggests that CEACAM41 is secreted by lymphoid cells upon activation. The results of our study demonstrate that alternative splicing after gene duplication is a potent mechanism to accelerate functional diversification of the equine CEA gene family members. This potent mechanism has created novel CEACAM receptors with unique signaling capacities and secreted CEACAMs which potentially enables equine lymphoid cells to control distantly located immune cells.
Get Full Text
Publication Date: 2018-03-15 PubMed ID: 29544443PubMed Central: PMC5856374DOI: 10.1186/s12862-018-1145-xGoogle Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research explores the evolution of the CEA gene family, specifically CEACAM1, in horses and its impact on the immune system. It suggests that gene duplication and alternative splicing are key mechanisms behind this gene family’s expansion and diversification.

Objective and Background

  • The researchers investigated the CEA gene family in the equine genome to understand how its expansion and diversification takes place. The CEA gene family, particularly its CEACAM1 component, is implicated in the immune system and shows varying degrees of expansion across different mammalian species. Underscoring this investigation is the observation that this gene family rapidly evolves in the human genome.

Research Findings

  • The team discovered that the horse genome contains 17 functional genes related to CEACAM1. Of these, nine were identified as secreted molecules and eight were CEACAMs comprising transmembrane and cytoplasmic domain exons.
  • A single gene, labeled as CEACAM41, was found to code for activating signaling motifs. The rest contained cytoplasmic exons similar to those of the inhibitory receptor CEACAM1.
  • Upon cloning cDNA, the researchers observed that only one CEACAM1 paralog contained functional immunoreceptor tyrosine-based inhibitory motifs in its cytoplasmic tail. Moreover, three receptors had acquired a stop codon in the transmembrane domain due to alternative splicing events, while two lost their inhibitory motifs.
  • Alternative splicing also transformed the function of an activating receptor into a secreted molecule by removing the transmembrane exon sequence.

Implications and Conclusion

  • A significant implication from these findings is that discovering the presence of these alternatively spliced CEACAMs in vivo suggests they can be expressed in living organisms. For instance, the detection of CEACAM41 mRNA in activated PBMC hints that this specific gene is secreted by lymphoid cells when activated.
  • This research proposes that alternative splicing following gene duplication is a significant driver of functional diversification within the equine CEA gene family. This method generated new CEACAM receptors with unique signaling abilities and secreted CEACAMs, thereby potentially allowing horse lymphoid cells to control distantly located immune cells.

Cite This Article

APA
Mißbach S, Aleksic D, Blaschke L, Hassemer T, Lee KJ, Mansfeld M, Hänske J, Handler J, Kammerer R. (2018). Alternative splicing after gene duplication drives CEACAM1-paralog diversification in the horse. BMC Evol Biol, 18(1), 32. https://doi.org/10.1186/s12862-018-1145-x

Publication

BMC evolutionary biology
ISSN: 1471-2148
NlmUniqueID: 100966975
Country: England
Language: English
Volume: 18
Issue: 1
Pages: 32
PII: 32

Researcher Affiliations

Mißbach, Sophie
  • Institute of Immunology, Friedrich-Loeffler-Institut, Suedufer 10, Greifswald, Insel Riems, Germany.
  • Plattform Degenerative Erkrankungen, Deutsches Primatenzentrum GmbH, Goettingen, Germany.
Aleksic, Denis
  • Institute of Immunology, Friedrich-Loeffler-Institut, Suedufer 10, Greifswald, Insel Riems, Germany.
Blaschke, Lisa
  • Institute of Immunology, Friedrich-Loeffler-Institut, Suedufer 10, Greifswald, Insel Riems, Germany.
Hassemer, Timm
  • Institute of Immunology, Friedrich-Loeffler-Institut, Suedufer 10, Greifswald, Insel Riems, Germany.
  • Department of Cellular Biochemistry, Max-Planck-Institute of Biochemistry, Martinsried, Germany.
Lee, Kyung Jin
  • Institute of Immunology, Friedrich-Loeffler-Institut, Suedufer 10, Greifswald, Insel Riems, Germany.
  • Department of Chemistry, Korea Advanced Institute of Science and Technology (KAIST), Daejeon, Republic of Korea.
Mansfeld, Martin
  • Institute of Immunology, Friedrich-Loeffler-Institut, Suedufer 10, Greifswald, Insel Riems, Germany.
Hänske, Jana
  • Institute of Immunology, Friedrich-Loeffler-Institut, Suedufer 10, Greifswald, Insel Riems, Germany.
Handler, Johannes
  • Clinic for Horses, Veterinary Faculty, Freie Universität Berlin, Oertzenweg 19b, D-14163, Berlin, Germany.
Kammerer, Robert
  • Institute of Immunology, Friedrich-Loeffler-Institut, Suedufer 10, Greifswald, Insel Riems, Germany. Robert.Kammerer@fli.bund.de.
  • Friedrich-Loeffler-Institut, Bundesforschungsinstitut für Tiergesundheit, Federal Research Institute for Animal Health, Südufer 10, D, 17493, Greifswald, Insel Riems, Germany. Robert.Kammerer@fli.bund.de.

MeSH Terms

  • Alternative Splicing / genetics
  • Amino Acid Motifs
  • Amino Acid Sequence
  • Animals
  • Antigens, CD / chemistry
  • Antigens, CD / genetics
  • Base Sequence
  • Cell Adhesion Molecules / chemistry
  • Cell Adhesion Molecules / genetics
  • Codon / genetics
  • Exons / genetics
  • Gene Duplication
  • Genetic Variation
  • Horses / genetics
  • Humans
  • Leukocytes, Mononuclear / metabolism
  • Protein Domains
  • Protein Isoforms / genetics
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Rabbits
  • Sequence Homology, Nucleic Acid

Grant Funding

  • (KF2875802UL2) / Bundesministerium fu00fcr Wirtschaft und Technologie
  • (Contract no. 81170269; Project No. 13.1432.7---001.00) / Gesellschaft fu00fcr internationale Zusammenarbeit
  • HE 6249/4-1 / Deutsche Forschungsgemeinschaft

Conflict of Interest Statement

ETHICS APPROVAL AND CONSENT TO PARTICIPATE: Healthy horses were slaughtered for meat production at the abattoir “Beerwart, Waiblingen”, not as part of this study, however we got permission from the abattoir to use of the tissues for the present study. Further tissue collection was approved by animal use committee of local authorities (Landesamt für Landwirtschaft, Lebensmittelsicherheit und Fischerei (LALLF) Rostock, Germany; 7221.3-2.1-011/13). CONSENT FOR PUBLICATION: Not applicable. COMPETING INTERESTS: The authors declare that they have no competing interests. PUBLISHER’S NOTE: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

References

This article includes 37 references
  1. Kammerer R, Hahn S, Singer BB, Luo JS, von Kleist S. Biliary glycoprotein (CD66a), a cell adhesion molecule of the immunoglobulin superfamily, on human lymphocytes: structure, expression and involvement in T cell activation.. Eur J Immunol 1998 Nov;28(11):3664-74.
    doi: 10.1002/(SICI)1521-4141(199811)28:11<3664::AID-IMMU3664>3.0.CO;2-Dpubmed: 9842909google scholar: lookup
  2. Kammerer R, Popp T, Singer BB, Schlender J, Zimmermann W. Identification of allelic variants of the bovine immune regulatory molecule CEACAM1 implies a pathogen-driven evolution.. Gene 2004 Sep 15;339:99-109.
    doi: 10.1016/j.gene.2004.06.023pubmed: 15363850google scholar: lookup
  3. Adler H, El-Gogo S, Guggemoos S, Zimmermann W, Beauchemin N, Kammerer R. Perturbation of lytic and latent gammaherpesvirus infection in the absence of the inhibitory receptor CEACAM1.. PLoS One 2009 Jul 21;4(7):e6317.
    doi: 10.1371/journal.pone.0006317pmc: PMC2708915pubmed: 19621080google scholar: lookup
  4. Adler H, Steer B, Juskewitz E, Kammerer R. To the editor: Murine gammaherpesvirus 68 (MHV-68) escapes from NK-cell-mediated immune surveillance by a CEACAM1-mediated immune evasion mechanism.. Eur J Immunol 2014 Aug;44(8):2521-2.
    doi: 10.1002/eji.201444593pubmed: 24976512google scholar: lookup
  5. Boulton IC, Gray-Owen SD. Neisserial binding to CEACAM1 arrests the activation and proliferation of CD4+ T lymphocytes.. Nat Immunol 2002 Mar;3(3):229-36.
    doi: 10.1038/ni769pubmed: 11850628google scholar: lookup
  6. Sintsova A, Sarantis H, Islam EA, Sun CX, Amin M, Chan CH, Stanners CP, Glogauer M, Gray-Owen SD. Global analysis of neutrophil responses to Neisseria gonorrhoeae reveals a self-propagating inflammatory program.. PLoS Pathog 2014 Sep;10(9):e1004341.
    doi: 10.1371/journal.ppat.1004341pmc: PMC4154863pubmed: 25188454google scholar: lookup
  7. Kammerer R, Zimmermann W. Coevolution of activating and inhibitory receptors within mammalian carcinoembryonic antigen families.. BMC Biol 2010 Feb 4;8:12.
    doi: 10.1186/1741-7007-8-12pmc: PMC2832619pubmed: 20132533google scholar: lookup
  8. Zimmermann W, Kammerer R. Coevolution of paired receptors in Xenopus carcinoembryonic antigen-related cell adhesion molecule families suggests appropriation as pathogen receptors.. BMC Genomics 2016 Nov 16;17(1):928.
    doi: 10.1186/s12864-016-3279-9pmc: PMC5112662pubmed: 27852220google scholar: lookup
  9. Kammerer R, Mansfeld M, Hänske J, Mißbach S, He X, Köllner B, Mouchantat S, Zimmermann W. Recent expansion and adaptive evolution of the carcinoembryonic antigen family in bats of the Yangochiroptera subgroup.. BMC Genomics 2017 Sep 11;18(1):717.
    doi: 10.1186/s12864-017-4106-7pmc: PMC5594555pubmed: 28893191google scholar: lookup
  10. Han E, Phan D, Lo P, Poy MN, Behringer R, Najjar SM, Lin SH. Differences in tissue-specific and embryonic expression of mouse Ceacam1 and Ceacam2 genes.. Biochem J 2001 Apr 15;355(Pt 2):417-23.
    doi: 10.1042/bj3550417pmc: PMC1221753pubmed: 11284729google scholar: lookup
  11. Thompson JA. Molecular cloning and expression of carcinoembryonic antigen gene family members.. Tumour Biol 1995;16(1):10-6.
    pubmed: 7863217doi: 10.1159/000217923google scholar: lookup
  12. Naghibalhossaini F, Stanners CP. Minimal mutations are required to effect a radical change in function in CEA family members of the Ig superfamily.. J Cell Sci 2004 Feb 15;117(Pt 5):761-9.
    doi: 10.1242/jcs.00903pubmed: 14734654google scholar: lookup
  13. Kammerer R, Popp T, Härtle S, Singer BB, Zimmermann W. Species-specific evolution of immune receptor tyrosine based activation motif-containing CEACAM1-related immune receptors in the dog.. BMC Evol Biol 2007 Oct 18;7:196.
    doi: 10.1186/1471-2148-7-196pmc: PMC2110893pubmed: 17945019google scholar: lookup
  14. Weichselbaumer M, Willmann M, Reifinger M, Singer J, Bajna E, Sobanov Y, Mechtcherikova D, Selzer E, Thalhammer JG, Kammerer R, Jensen-Jarolim E. Phylogenetic discordance of human and canine carcinoembryonic antigen (CEA, CEACAM) families, but striking identity of the CEA receptors will impact comparative oncology studies.. PLoS Curr 2011 Mar 16;3:RRN1223.
    doi: 10.1371/currents.RRN1223pmc: PMC3059814pubmed: 21436956google scholar: lookup
  15. Aleksic D, Blaschke L, Mißbach S, Hänske J, Weiß W, Handler J, Zimmermann W, Cabrera-Sharp V, Read JE, de Mestre AM, O'Riordan R, Moore T, Kammerer R. Convergent evolution of pregnancy-specific glycoproteins in human and horse.. Reproduction 2016 Sep;152(3):171-84.
    doi: 10.1530/REP-16-0236pubmed: 27280409google scholar: lookup
  16. Moore T, Dveksler GS. Pregnancy-specific glycoproteins: complex gene families regulating maternal-fetal interactions.. Int J Dev Biol 2014;58(2-4):273-80.
    doi: 10.1387/ijdb.130329gdpubmed: 25023693google scholar: lookup
  17. Pavlopoulou A, Scorilas A. A comprehensive phylogenetic and structural analysis of the carcinoembryonic antigen (CEA) gene family.. Genome Biol Evol 2014 May 23;6(6):1314-26.
    doi: 10.1093/gbe/evu103pmc: PMC4079198pubmed: 24858421google scholar: lookup
  18. Delgado Tascón J, Adrian J, Kopp K, Scholz P, Tschan MP, Kuespert K, Hauck CR. The granulocyte orphan receptor CEACAM4 is able to trigger phagocytosis of bacteria.. J Leukoc Biol 2015 Mar;97(3):521-31.
    doi: 10.1189/jlb.2AB0813-449RRpmc: PMC5477890pubmed: 25567962google scholar: lookup
  19. Nagel G, Grunert F, Kuijpers TW, Watt SM, Thompson J, Zimmermann W. Genomic organization, splice variants and expression of CGM1, a CD66-related member of the carcinoembryonic antigen gene family.. Eur J Biochem 1993 May 15;214(1):27-35.
    doi: 10.1111/j.1432-1033.1993.tb17892.xpubmed: 8508798google scholar: lookup
  20. Stoffel A, Neumaier M, Gaida FJ, Fenger U, Drzeniek Z, Haubeck HD, Wagener C. Monoclonal, anti-domain and anti-peptide antibodies assign the molecular weight 160,000 granulocyte membrane antigen of the CD66 cluster to a mRNA species encoded by the biliary glycoprotein gene, a member of the carcinoembryonic antigen gene family.. J Immunol 1993 Jun 1;150(11):4978-84.
    pubmed: 8496599
  21. Pan Q, Shai O, Lee LJ, Frey BJ, Blencowe BJ. Deep surveying of alternative splicing complexity in the human transcriptome by high-throughput sequencing.. Nat Genet 2008 Dec;40(12):1413-5.
    doi: 10.1038/ng.259pubmed: 18978789google scholar: lookup
  22. Barnett TR, Drake L, Pickle W 2nd. Human biliary glycoprotein gene: characterization of a family of novel alternatively spliced RNAs and their expressed proteins.. Mol Cell Biol 1993 Feb;13(2):1273-82.
    doi: 10.1128/MCB.13.2.1273pmc: PMC359012pubmed: 8423792google scholar: lookup
  23. Barnett TR, Kretschmer A, Austen DA, Goebel SJ, Hart JT, Elting JJ, Kamarck ME. Carcinoembryonic antigens: alternative splicing accounts for the multiple mRNAs that code for novel members of the carcinoembryonic antigen family.. J Cell Biol 1989 Feb;108(2):267-76.
    doi: 10.1083/jcb.108.2.267pmc: PMC2115422pubmed: 2537311google scholar: lookup
  24. Iñiguez LP, Hernández G. The Evolutionary Relationship between Alternative Splicing and Gene Duplication.. Front Genet 2017;8:14.
    doi: 10.3389/fgene.2017.00014pmc: PMC5306129pubmed: 28261262google scholar: lookup
  25. Eshel D, Toporik A, Efrati T, Nakav S, Chen A, Douvdevani A. Characterization of natural human antagonistic soluble CD40 isoforms produced through alternative splicing.. Mol Immunol 2008 Dec;46(2):250-7.
    doi: 10.1016/j.molimm.2008.08.280pubmed: 18849075google scholar: lookup
  26. Dery KJ, Kujawski M, Grunert D, Wu X, Ngyuen T, Cheung C, Yim JH, Shively JE. IRF-1 regulates alternative mRNA splicing of carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) in breast epithelial cells generating an immunoreceptor tyrosine-based inhibition motif (ITIM) containing isoform.. Mol Cancer 2014 Mar 21;13:64.
    doi: 10.1186/1476-4598-13-64pmc: PMC4113144pubmed: 24650050google scholar: lookup
  27. Nédellec P, Dveksler GS, Daniels E, Turbide C, Chow B, Basile AA, Holmes KV, Beauchemin N. Bgp2, a new member of the carcinoembryonic antigen-related gene family, encodes an alternative receptor for mouse hepatitis viruses.. J Virol 1994 Jul;68(7):4525-37.
    pmc: PMC236379pubmed: 8207827doi: 10.1128/JVI.68.7.4525-4537.1994google scholar: lookup
  28. Sapoznik S, Hammer O, Ortenberg R, Besser MJ, Ben-Moshe T, Schachter J, Markel G. Novel anti-melanoma immunotherapies: disarming tumor escape mechanisms.. Clin Dev Immunol 2012;2012:818214.
    doi: 10.1155/2012/818214pmc: PMC3386565pubmed: 22778766google scholar: lookup
  29. Barclay AN, Hatherley D. The counterbalance theory for evolution and function of paired receptors.. Immunity 2008 Nov 14;29(5):675-8.
    doi: 10.1016/j.immuni.2008.10.004pmc: PMC3128988pubmed: 19006692google scholar: lookup
  30. van Beek EM, Cochrane F, Barclay AN, van den Berg TK. Signal regulatory proteins in the immune system.. J Immunol 2005 Dec 15;175(12):7781-7.
    pubmed: 16339510doi: 10.4049/jimmunol.175.12.7781google scholar: lookup
  31. Barclay AN, Van den Berg TK. The interaction between signal regulatory protein alpha (SIRPα) and CD47: structure, function, and therapeutic target.. Annu Rev Immunol 2014;32:25-50.
    doi: 10.1146/annurev-immunol-032713-120142pubmed: 24215318google scholar: lookup
  32. Zebhauser R, Kammerer R, Eisenried A, McLellan A, Moore T, Zimmermann W. Identification of a novel group of evolutionarily conserved members within the rapidly diverging murine Cea family.. Genomics 2005 Nov;86(5):566-80.
    doi: 10.1016/j.ygeno.2005.07.008pubmed: 16139472google scholar: lookup
  33. Khairnar V, Duhan V, Maney SK, Honke N, Shaabani N, Pandyra AA, Seifert M, Pozdeev V, Xu HC, Sharma P, Baldin F, Marquardsen F, Merches K, Lang E, Kirschning C, Westendorf AM, Häussinger D, Lang F, Dittmer U, Küppers R, Recher M, Hardt C, Scheffrahn I, Beauchemin N, Göthert JR, Singer BB, Lang PA, Lang KS. CEACAM1 induces B-cell survival and is essential for protective antiviral antibody production.. Nat Commun 2015 Feb 18;6:6217.
    doi: 10.1038/ncomms7217pmc: PMC4346637pubmed: 25692415google scholar: lookup
  34. Kammerer R, Stober D, Singer BB, Obrink B, Reimann J. Carcinoembryonic antigen-related cell adhesion molecule 1 on murine dendritic cells is a potent regulator of T cell stimulation.. J Immunol 2001 Jun 1;166(11):6537-44.
    doi: 10.4049/jimmunol.166.11.6537pubmed: 11359805google scholar: lookup
  35. Jiang L, Barclay AN. Identification of leucocyte surface protein interactions by high-throughput screening with multivalent reagents.. Immunology 2010 Jan;129(1):55-61.
    doi: 10.1111/j.1365-2567.2009.03153.xpmc: PMC2807486pubmed: 20050330google scholar: lookup
  36. Singer BB, Opp L, Heinrich A, Schreiber F, Binding-Liermann R, Berrocal-Almanza LC, Heyl KA, Müller MM, Weimann A, Zweigner J, Slevogt H. Soluble CEACAM8 interacts with CEACAM1 inhibiting TLR2-triggered immune responses.. PLoS One 2014;9(4):e94106.
    doi: 10.1371/journal.pone.0094106pmc: PMC3990526pubmed: 24743304google scholar: lookup
  37. Markel G, Achdout H, Katz G, Ling KL, Salio M, Gruda R, Gazit R, Mizrahi S, Hanna J, Gonen-Gross T, Arnon TI, Lieberman N, Stren N, Nachmias B, Blumberg RS, Steuer G, Blau H, Cerundolo V, Mussaffi H, Mandelboim O. Biological function of the soluble CEACAM1 protein and implications in TAP2-deficient patients.. Eur J Immunol 2004 Aug;34(8):2138-48.
    doi: 10.1002/eji.200425021pubmed: 15259011google scholar: lookup

Citations

This article has been cited 4 times.
  1. Bonsignore P, Kuiper JWP, Adrian J, Goob G, Hauck CR. CEACAM3-A Prim(at)e Invention for Opsonin-Independent Phagocytosis of Bacteria. Front Immunol 2019;10:3160.
    doi: 10.3389/fimmu.2019.03160pubmed: 32117212google scholar: lookup
  2. Hänske J, Hammacher T, Grenkowitz F, Mansfeld M, Dau TH, Maksimov P, Friedrich C, Zimmermann W, Kammerer R. Natural selection supports escape from concerted evolution of a recently duplicated CEACAM1 paralog in the ruminant CEA gene family. Sci Rep 2020 Feb 25;10(1):3404.
    doi: 10.1038/s41598-020-60425-4pubmed: 32099040google scholar: lookup
  3. Kim SW, Jo A, Im J, Lee HE, Kim HS. Expression analysis of miR-221-3p and its target genes in horses. Genes Genomics 2019 Apr;41(4):459-465.
    doi: 10.1007/s13258-018-00778-3pubmed: 30604147google scholar: lookup
  4. Kammerer R, Ballesteros A, Bonsor D, Warren J, Williams JM, Moore T, Dveksler G. Equine pregnancy-specific glycoprotein CEACAM49 secreted by endometrial cup cells activates TGFB. Reproduction 2020 Nov;160(5):685-694.
    doi: 10.1530/REP-20-0277pubmed: 33065543google scholar: lookup
Subscribe to our newsletter
Join 99,075 horse owners like you who receive our email updates on horse health and nutrition.